Isoquinoline derivatives

ABSTRACT

The invention relates to 6-piperidinyl-substituted isoquinoline derivatives of the formula (I) 
                         
useful for the treatment and/or prevention of diseases associated with Rho-kinase and/or Rho-kinase mediated phosphorylation of myosin light chain phosphatase, and compositions containing such compounds.

This application is a Divisional of U.S. patent application Ser. No.11/961,193 filed Dec. 20, 2007, which is a Continuation of InternationalApplication No. PCT/EP2006/005648, filed Jun. 13, 2006, both of whichare incorporated herein by reference in their entirety.

The present invention relates to novel isoquinoline derivatives, theirpreparation and their use in the treatment and/or prevention of diseasesrelated to the inhibition of Rho-kinase and/or of Rho-kinase mediatedphosphorylation of myosin light chain phosphatase.

Activation of a small GTPase RhoA upon agonist stimulation results inconversion of RhoA from the inactive GDP-bound form to the activeGTP-bound form with a subsequent binding to and activation ofRho-kinase. Two isoforms, Rho-kinase 1 and Rho-kinase 2, are known.Rho-kinase 2 is expressed in vascular smooth muscle cells andendothelial cells. Activation of Rho-kinase 2 by the active GTP-boundRhoA leads to calcium sensitization of smooth muscle cells throughphosphorylation-mediated inhibition of the myosin light chainphosphatase activity and thereby up-regulation of the activity of myosinregulatory light chain (Uehata et al., Nature 1997, 389, 990-994).

It is known that Rho-kinase is involved in vasoconstriction, includingthe development of myogenic tone (J. Appl. Physiol. 2005, 1940-8, 98),bronchial smooth muscle contraction (Am. J. Resp. Cell Mol. Biol. 20,1190-1200), hypertension, i.e. pulmonary hypertension (Heart, 91, 391-2,2005) and ocular hypertension (Invest. Ophthalmol. Visual Sci. 2001, 42,137-144), endothelial dysfunction (Eur. J. Pharmacol. 2005, 512,247-249), artherosclerosis, restenosis (Arch. Mal. Coeur 2005, 98,249-254), glucose utilization, cardiac hypertrophy (Hypertension 2000,35, 313-318), erectile dysfunction (Nature Medicine 2001, 7, 119-122),retinopathy, inflammation, immune diseases, AIDS, osteoporosis, brainfunctional disorder, infection of digestive tracts with bacteria (WO98/06433), cancer development, vascular smooth muscle proliferation andmotility (Circ. Res. 1999, 84, 1186-1193; Atherosclerosis 2001, 155,321-327), endothelial proliferation and motility (Biochem. Biophys. Res.Commun. 2000, 269, 633-640), stress fiber formation (Science 1997, 275,1308-1311; J. Cell Biol. 2000, 150, 797-806), platelet aggregation (FEBSLett. 2000, 466, 70-74; Blood 1999, 94, 1665-1672), Na/H exchangetransport system activation (EMBO J. 1998, 17, 4712-4722), Alzheimer'sdisease (Science 2003, 302, 1215-1217), adducin activation (J. Biol.Chem., 273, 5542-5548, 1998), and in SREB (Sterol response bindingelement) signalling and its effects on lipid metabolism (Circ. Res., 92,1296-304, 2003).

Therefore, a compound having inhibitory effect on Rho-kinase and/or onRho-kinase mediated phosphorylation of myosin light chain phosphatase isuseful for the treatment and/or prevention of diseases involvingRho-kinase as the primary disease cause, e.g. hypertension, i.e.,pulmonary hypertension and ocular hypertension, peripheral circulatorydisorder, angina pectoris, cerebral vasospasm, asthma, premature birth,hyperaggregability of platelets, Peripheral Occlusive Arterial Disease(PAOD), Chronic Obstructive Pulmonary Disease (COPD), cancerdevelopment, and erectile dysfunction, or as the secondary diseasecause, e.g. arteriosclerosis, ischemic organ failure (end organ damage),fibroid lung, fibroid liver, liver failure, fibroid kidney, renalglomerulosclerosis, kidney failure, organ hypertrophy, prostatichypertrophy, complications of diabetes, blood vessel restenosis,atherosclerosis, cancer, cardiac hypertrophy, heart failure; ischemicdiseases; inflammation; autoimmune diseases; AIDS, osteopathy such asosteoporosis, brain functional disorder, infection of digestive tractswith bacteria, sepsis, adult respiratory distress syndrome, retinopathy,glaucoma and Alzheimer's disease.

WO 01/64238 describes isoquinoline-5-sulfonamide derivatives optionallysubstituted by a —(CH₂)₁₋₆—O—(CH₂)₀₋₆—, a —(CH₂)₀₋₆—S—(CH₂)₀₋₆— or a—(CH₂)₀₋₆-linked heterocyclic group useful as neuroprotective agents.

JP 10087629 A describes isoquinoline derivatives useful for thetreatment of infections caused by Heliobacter pylori such as for examplegastritis or ulcer. The isoquinoline derivatives are preferably5-substituted by X—[(C₁-C₆)alkylene)]₀₋₁-Y wherein X may be oxygen and Ymay be an aryl or a heterocyclic group.

Hagihara et al. (Bioorg. Med. Chem. 1999, 7, 2647-2666) disclose6-benzyloxy-isoquinoline for the treatment of infections caused byHeliobacter pylori.

U.S. Pat. No. 5,480,883 generically discloses as EGF and/or PDGFreceptor inhibitors useful for inhibiting cell proliferation compoundsof the formula “Ar I-X-Ar II” wherein X may be (CHR₁)_(m)—Z—(CHR₁)_(n),e.g. Z—CH₂, wherein Z may be O, R₁ is hydrogen or alkyl, Ar I may beamong others an optionally substituted C₅₋₁₂ bicyclic heteroaryl ringsystem and Ar II may be among others an optionally substituted C₃₋₇monocyclic saturated heterocyclic system.

WO 03/053330 describes isoquinoline derivatives of the formula

wherein L² is halogen and R¹⁰ may be C₁₋₅alkylene-C₅₋₆heterocyclic groupas intermediates in the synthesis of GSK-3 inhibitors.

An embodiment of the present invention is a compound of the formula (I)

whereinR₁ isH,(C₁-C₆)alkyl,R′,NH—(C₁-C₆)alkyl,NH—R′, orN[(C₁-C₆)alkyl]₂;R₂ is hydrogen, halogen, or (C₁-C₆)alkyl;R₃ isH,halogen,(C₁-C₆)alkyl,(C₁-C₆)alkylene-R′,OH,O—R″,NH₂,NHR″,NR″R″ orNH—C(O)—R″,R₄ isH,halogen,hydroxy,CN,(C₁-C₆)alkyl,R′,(C₁-C₆)alkylene-R′;R₅ isH,halogen,CN,NO₂,(C₁-C₆)alkyl,(C₂-C₆)alkenyl,R′,(C₁-C₆)alkylene-(C₆-C₁₀)aryl,(C₂-C₆)alkenylene-(C₆-C₁₀)aryl,(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl,CH(OH)—(C₁-C₆)alkyl,NH₂,NH—R′,NH—SO₂H,NH—SO₂—(C₁-C₆)alkyl,NH—SO₂—R′,NH—C(O)—(C₁-C₆)alkyl,NH—C(O)—R′,C(O)N[(C₁-C₆)alkyl]₂,C(O)OH, orC(O)O—(C₁-C₆)alkyl;R₆ isH,R′,(C₁-C₈)alkyl,(C₁-C₆)alkylene-R′,(C₁-C₆)alkylene-O—(C₁-C₆)alkyl,(C₁-C₆)alkylene-O—R′,(C₁-C₆)alkylene-CH[R′]₂,(C₁-C₆)alkylene-C(O)—R′,(C₁-C₆)alkylene-C(O)NH₂,(C₁-C₆)alkylene-C(O)NH—R′, or(C₁-C₆)alkylene-C(O)N[R′]₂;R₇ isH,halogen,CN,NO₂,(C₁-C₆)alkyl,(C₂-C₆)alkenyl,R′,(C₂-C₆)alkenylene-(C₆-C₁₀)aryl,(C₁-C₆)alkylene-R′,CH(OH)—(C₁-C₆)alkyl,CH(OH)—(C₆-C₁₀)aryl,CH(OH)—(C₅-C₁₀)heterocyclyl,NH₂,NH—R′,NH—SO₂H,NH—SO₂—(C₁-C₆)alkyl,NH—SO₂—R′,SO₂—NH₂,SO₂—NHR′,NH—C(O)—(C₁-C₆)alkyl,NH—C(O)—R′,C(O)N[(C₁-C₆)alkyl]₂,C(O)OH, orC(O)O—(C₁-C₆)alkyl;R₈ is H, halogen or (C₁-C₆)alkyl;n is 1, 2, 3 or 4;L is O or O—(C₁-C₆)alkylene;R′ is(C₃-C₈)cycloalkyl,(C₅-C₁₀)heterocyclyl,(C₆-C₁₀)aryl; andR″ is(C₃-C₈)cycloalkyl,(C₅-C₁₀)heterocyclyl,(C₆-C₁₀)aryl,(C₁-C₆)alkyl,(C₁-C₆)alkylene-R′,(C₁-C₆)alkylene-O—(C₁-C₆)alkyl,(C₁-C₆)alkylene-O—R′, or(C₁-C₆)alkylene-NR_(x)R_(y); andR_(x) and R_(y) are independently of each other(C₁-C₆)alkyl,(C₅-C₁₀)heterocyclyl,(C₆-C₁₀)aryl,(C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-(C₆-C₁₀)aryl,(C₁-C₄)alkylene-NH(C₁-C₆)alkyl,(C₁-C₄)alkylene-N[(C₁-C₆)alkyl]₂,(C₁-C₄)alkylene-N[(C₆-C₁₀)aryl]₂, or(C₁-C₄)alkylene-N[(C₅-C₁₀)heterocyclyl]₂; andwherein in residues R₄, R₅, R₇ and R₈ one alkyl or alkylene hydrogenatom can optionally be substituted by OH, F, OCH₃, COOH, COOCH₃, NH₂,NHCH₃, N(CH₃)₂, CONH₂, CONHCH₃ or CON(CH₃)₂;or a pharmaceutically acceptable salt thereof and/or a physiologicallyfunctional derivative thereof.

Preferably, R₁ is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, NH—(C₁-C₆)alkyl,NH—(C₆-C₁₀)aryl or N[(C₁-C₆)alkyl]₂. More preferably, R₁ is H, halogen,(C₁-C₄)alkyl, NH—(C₁-C₄)alkyl, N[(C₁-C₄)alkyl]₂ or NH-phenyl. Mostpreferably, R₁ is H, (C₁-C₂)alkyl or NH—(C₁-C₂)alkyl, especiallypreferred H.

Preferably, R₂ is H, halogen or (C₁-C₄)alkyl. Preferably, R₂ is H or(C₁-C₄)alkyl. More preferred, R₂ is H, (C₁-C₂)alkyl. R₂ may be bound toany carbon atom of the piperidine ring including the position where thelinker group L is bound.

R₃ is preferably H, halogen, (C₁-C₄)alkylene-R′, O—R″ or NHR″. Morepreferred, R₃ is H or NHR″. Most preferred, R₃ is H,NH—(C₅-C₆)heterocyclyl or NH-phenyl, especially preferred are H,NH—(C₅-C₆)heteroaryl containing one or more N atoms or NH-phenyl. Mostespecially preferred, R₃ is H. Examples of R₃ substituents are

Preferably, R₄ is H, halogen or (C₁-C₆)alkyl. More preferred, R₄ is H,halogen or (C₁-C₄)alkyl. Most preferred, R₄ is H.

Preferably, R₅ is H, halogen, CN, (C₁-C₆)alkyl, R′, NH—(C₆-C₁₀)aryl or(C₁-C₆)alkylene-R′. More preferably, R₅ is H, halogen, (C₁-C₆)alkyl, R′,NH—(C₆-C₁₀)aryl or (C₁-C₆)alkylene-R′. Most preferably, R₅ is H,halogen, (C₆-C₁₀)aryl, NH—(C₆-C₁₀)aryl, (C₁-C₂)alkyl-(C₆-C₁₀)aryl,(C₁-C₆)alkyl or (C₅-C₁₀)heteroaryl. Especially preferred, R₅ is H,halogen, phenyl, (C₁-C₆)alkyl or (C₅-C₆)heteroaryl. Examples of R₅ arehydrogen, fluoro, chloro, bromo, iodo, nitrile, nitro,(p-methoxy)-phenyl, N-aniline, phenyl, benzyl, methyl, ethyl, vinyl,2-propenyl, s-butenyl, cyclopropyl, thienyl, tetrazol, amino,4-methoxy-anilin, N-acetyl or a substituent of the group consisting of

Preferably, R₆ is H, (C₁-C₆)alkyl, R′,(C₁-C₄)alkylene-(C₃-C₈)cycloalkyl, (C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-C(O)—(C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-C(O)—(C₆-C₁₀)aryl or (C₁-C₆)alkylene-(C₆-C₁₀)aryl. Morepreferred, R₆ is H, (C₁-C₆)alkyl, (C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl or (C₁-C₆)alkylene-(C₆-C₁₀)aryl.Examples of R₆ are H, methyl, ethyl, propyl, butyl, s-butyl, pentyl,3-methyl-butyl, isopropyl, trifluoromethyl, 3,3,3-trifluorobutyl,cyclopropyl, methylene cyclopropyl, 2-pyrimidinyl, benzyl or asubstituent of the group consisting of

Preferably, R₇ is H, halogen, CN, (C₁-C₆)alkyl, (C₂-C₆)alkenyl, R′ or(C₁-C₆)alkylene-(C₃-C₈)cycloalkyl. More preferred, R₇ is H, halogen, CN,(C₁-C₄)alkyl, (C₁-C₄)alkenyl, phenyl, cyclopropyl or (C₅-C₆)heteroaryl.Most preferably, R₇ is H, fluoro, chloro, bromo, methyl, ethyl, phenyl,nitrile, cyclopropyl, thienyl or vinyl.

R₈ is preferably H, halogen or (C₁-C₄)alkyl. More preferred, R₈ is H,Cl, F, methyl or ethyl.

Preferably, n is 1, 2 or 3. More preferred, n is 1.

The linker group L may be bound to the piperidine ring in any positionvia a piperidine ring carbon atom. In a preferred embodiment, L isattached to the 4-position of the piperidine ring

L is attached to the 3-position of the piperidine ring

In an especially preferred embodiment, L is attached to the 4-positionof the piperidine ring.

In a further preferred embodiment, L is O-methylene, O-ethylene orpreferably O. More preferably, L is O-methylene, O-ethylene or Oattached to the 4-position of the piperidine ring.

In preferred embodiments of the present invention one or more or all ofthe groups contained in the compounds of formula (I) can independentlyof each other have any of the preferred, more preferred or mostpreferred definitions of the groups specified above or any one or someof the specific denotations which are comprised by the definitions ofthe groups and specified above, all combinations of preferreddefinitions, more preferred or most preferred and/or specificdenotations being a subject of the present invention. Also with respectto all preferred embodiments the invention includes the compounds of theformula (I) in all stereoisomeric forms and mixtures of stereoisomericforms in all ratios, and their physiologically acceptable salts.

The term “*-” in the exemplified substituents vide supra marks the pointwhere the substituent is attached, which means, for example, for a R₃substituent

a compound of the formula

A preferred embodiment is a compound of the formula (I) wherein

R₁ is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, NH—(C₁-C₆)alkyl, NH—(C₆-C₁₀)aryl,or N[(C₁-C₆)alkyl]₂;

R₂ is hydrogen, halogen, or (C₁-C₆)alkyl;

R₃ is H, halogen, (C₁-C₄)alkylene-R′, O—R″ or NHR″, wherein R′ and R″are defined as above;

R₄ is H, halogen or (C₁-C₆)alkyl;

R₅ is H, halogen, (C₁-C₆)alkyl, CN, (C₆-C₁₀)aryl, NH—(C₆-C₁₀)aryl,(C₁-C₆)alkylene-(C₆-C₁₀)aryl, (C₅-C₁₀)heterocyclyl or(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl;

R₆ is H, R′, (C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl,(C₁-C₆)alkylene-C(O)—(C₆-C₁₀)aryl,(C₁-C₄)alkylene-C(O)—(C₅-C₁₀)heterocyclyl, (C₁-C₆)alkylene-(C₆-C₁₀)arylor (C₁-C₆)alkyl.

R₇ is H, halogen, CN, (C₁-C₆)alkyl, (C₂-C₆)alkenyl or R′;

R₈ is H, halogen or (C₁-C₆)alkyl;

n is 1, 2 or 3, and

L is O, O-methylene or O-ethylene;

or a pharmaceutically acceptable salt thereof and/or a physiologicallyfunctional derivative thereof.

A further preferred embodiment is a compound of the formula (I) wherein

R₁ is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, NH—(C₁-C₆)alkyl, NH—(C₆-C₁₀)aryl,or N[(C₁-C₆)alkyl]₂;

R₂ is H or (C₁-C₄)alkyl;

R₃ is H, halogen or NHR″, wherein R″ is defined as above;

R₄ is H, halogen or (C₁-C₄)alkyl;

R₅ is H, halogen, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, NH—(C₆-C₁₀)aryl,(C₁-C₆)alkylene-(C₆-C₁₀)aryl or (C₅-C₁₀)heterocyclyl;

R₆ is H, (C₁-C₆)alkyl, R′, (C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl or(C₁-C₆)alkylene-(C₆-C₁₀)aryl;

R₇ is H, halogen, CN, (C₁-C₆)alkyl, (C₂-C₆)alkenyl or R′;

R₈ is H, halogen or (C₁-C₆)alkyl;

n is 1, 2 or 3; and

L is O;

or a pharmaceutically acceptable salt thereof and/or a physiologicallyfunctional derivative thereof.

An especially preferred embodiment is a compound of the formula (I)wherein

R₁ is H, (C₁-C₄)alkyl, NH—(C₁-C₄)alkyl, N[(C₁-C₄)alkyl]₂ or NH-phenyl;

R₂ is H, (C₁-C₄)alkyl;

R₃ is H, NH—(C₅-C₆)heteroaryl or NH-phenyl;

R₄ is H, halogen or (C₁-C₄)alkyl;

R₅ is H, halogen, (C₁-C₄)alkyl, (C₆-C₁₀)aryl, NH—(C₆-C₁₀)aryl,(C₁-C₂)alkyl-(C₆-C₁₀)aryl or (C₅-C₁₀)heteroaryl;

R₆ is H, (C₁-C₆)alkyl, (C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-(C₅-C₁₀)heterocyclyl, (C₆-C₁₀)aryl or(C₁-C₆)alkylene-(C₆-C₁₀)aryl;

R₇ is H, halogen, CN, (C₁-C₄)alkyl, (C₁-C₄)alkenyl, phenyl, cyclopropyl,(C₅-C₆)heteroaryl;

R₈ is H, halogen or (C₁-C₄)alkyl;

n is 1; and

L is O;

or a pharmaceutically acceptable salt thereof and/or a physiologicallyfunctional derivative thereof.

As in any embodiment of the invention, in the preceding embodimentswhich contain preferred, more preferred, most preferred or exemplarydefinitions of compounds according to the invention, one or more or allof the groups can have any of its preferred, more preferred, mostpreferred definitions specified above or any one or some of the specificdenotations which are comprised by its definitions and are specifiedabove.

Isoquinoline and piperidyl substitution pattern are numbered textaccording to IUPAC rules:

Physiologically acceptable salts of compounds of the formula (I) meanboth their organic and inorganic salts as described in Remington'sPharmaceutical Sciences (17th edition, page 1418 (1985)). Because of thephysical and chemical stability and the solubility, preference is givenfor acidic groups inter alia to sodium, potassium, calcium and ammoniumsalts; preference is given for basic groups inter alia to salts ofmaleic acid, fumaric acid, succinic acid, malic acid, tartaric acid,methylsulfonic acid, hydrochloric acid, sulfuric acid, phosphoric acidor of carboxylic acids or sulfonic acids, for example as hydrochlorides,hydrobromides, phosphates, sulfates, methanesulfonates, acetates,lactates, maleates, fumarates, malates, gluconates, and salts of aminoacids, of natural bases or carboxylic acids. The preparation ofphysiologically acceptable salts from compounds of the formula (I) and(II) which are capable of salt formation, including their stereoisomericforms, takes place in a manner known per se. The compounds of theformula (I) form stable alkali metal, alkaline earth metal or optionallysubstituted ammonium salts with basic reagents such as hydroxides,carbonates, bicarbonates, alcoholates and ammonia or organic bases, forexample trimethyl- or triethylamine, ethanolamine, diethanolamine ortriethanolamine, trometamol or else basic amino acids, for examplelysine, ornithine or arginine. Where the compounds of the formula (I)have basic groups, stable acid addition salts can also be prepared withstrong acids. Suitable pharmaceutically acceptable acid addition saltsof the compounds of the invention are salts of inorganic acids such ashydrochloric acid, hydrobromic, phosphoric, metaphosphoric, nitric andsulfuric acid, and of organic acids such as, for example, acetic acid,benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic,glycolic, isethionic, lactic, lactobionic, maleic, malic,methanesulfonic, succinic, p-toluenesulfonic and tartaric acid.

Salts with a physiologically unacceptable anion such as, for example,trifluoroacetate likewise belong within the framework of the inventionas useful intermediates for the preparation or purification ofpharmaceutically acceptable salts and/or for use in nontherapeutic, forexample in vitro, applications.

The term “physiologically functional derivative” used herein refers toany physiologically tolerated derivative of a compound of the formula(I) of the invention, for example an N-oxide, which on administration toa mammal such as, for example, a human is able to form (directly orindirectly) a compound of the formula (I) or an active metabolitethereof.

Physiologically functional derivatives include prodrugs of the compoundsof the invention, as described, for example, in H. Okada et al., Chem.Pharm. Bull. 1994, 42, 57-61. Such prodrugs can be metabolized in vivoto a compound of the invention. These prodrugs may themselves be activeor not.

The invention relates to compounds of the formula (I) in the form oftheir racemates, racemic mixtures and pure enantiomers and to theirdiastereomers and mixtures thereof.

If radicals or substituents may occur more than once in the compounds ofthe formula (I), they may all, independently of one another, have thestated meaning and be identical or different.

The compounds of the invention may also exist in various polymorphousforms, for example as amorphous and crystalline polymorphous forms. Allpolymorphous forms of the compounds of the invention belong within theframework of the invention and are a further aspect of the invention.

All references to “compound(s) of formula (I)” hereinafter refer tocompound(s) of the formula (I) as described above, and theirphysiologically acceptable salts, solvates and physiologicallyfunctional derivatives as described herein.

The terms (C₁-C₂)alkyl, (C₁-C₄)alkyl, (C₁-C₆)alkyl, (C₁-C₈)alkyl and thecorresponding alkylene substituents are understood as a hydrocarbonresidue which can be linear, i.e. straight-chain, or branched and has 1,2, 3, 4, 5, 6, 7 or 8 carbon atoms, respectively. This also applies ifan alkyl group occurs as a substituent on another group, for example inan alkoxy group (O-alkyl), S-alkyl or a —O(C₁-C₆)alkylene-O—, analkoxycarbonyl group or an arylalkyl group. Examples of alkyl groups aremethyl, ethyl, propyl, butyl, pentyl or hexyl, the n-isomers of allthese groups, isopropyl, isobutyl, 1-methylbutyl, isopentyl, neopentyl,2,2-dimethylbutyl, 2-methylpentyl, 3-methylpentyl, isohexyl, sec-butyl,tert-butyl or tert-pentyl. Alkyl groups may—if not otherwise stated—behalogenated once or more, i.e. alkyl groups may be fluorinated, i.e.perfluorinated. Examples of halogenated alkyl groups are CF₃ and CH₂CF₃,OCF₃, SCF₃, or —O—(CF₂)₂—O—.

Alkenyl are, for example, vinyl, 1-propenyl, 2-propenyl (=allyl),2-butenyl, 3-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 5-hexenylor 1,3-pentadienyl.

Alkynyl are, for example, ethynyl, 1-propynyl, 2-propynyl (=propargyl)or 2-butynyl.

Halogen means fluoro, chloro, bromo or iodo.

(C₃-C₈)cycloalkyl groups are cyclic alkyl groups containing 3, 4, 5, 6,7 or 8 ring carbon atoms like cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl or cyclooctyl, which can also be substituted and/or contain 1or 2 double bounds (unsaturated cycloalkyl groups) like, for example,cyclopentenyl or cyclohexenyl can be bonded via any carbon atom.

A (C₆-C₁₀)aryl group means an aromatic ring or a ring system whichcomprises two aromatic rings which are fused or otherwise linked, forexample a phenyl, naphthyl, biphenyl, tetrahydronaphthyl, alpha- orbeta-tetralon-, indanyl- or indan-1-on-yl group. A preferred(C₆-C₁₀)aryl group is phenyl.

A (C₅-C₁₀)heterocyclyl group means a mono- or bicyclic ring system whichcomprises, apart from carbon, one or more heteroatoms such as, forexample, e.g. 1, 2 or 3 nitrogen atoms, 1 or 2 oxygen atoms, 1 or 2sulfur atoms or combinations of different hetero atoms. The heterocyclylresidues can be bound at any positions, for example on the 1-position,2-position, 3-position, 4-position, 5-position, 6-position, 7-positionor 8-position. (C₅-C₁₀)heterocyclyl groups may be (1) aromatic (i.e.,heteroaryl groups) or (2) saturated or (3) mixed aromatic/saturated.

Suitable (C₅-C₁₀)heterocyclyl group include acridinyl, azocinyl,benzimidazolyl, benzofuryl, benzomorpholinyl, benzothienyl,benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, carbazolyl,4aH-carbazolyl, carbolinyl, furanyl, quinazolinyl, quinolinyl,4H-quinolizinyl, quinoxalinyl, quinuclidinyl, chromanyl, chromenyl,chromen-2-onyl, cinnolinyl, decahydroquinolinyl,2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]-tetrahydrofuran, furyl,furazanyl, homomorpholinyl, homopiperazinyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl,3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl,isoindolyl, isoquinolinyl (benzimidazolyl), isothiazolyl, isoxazolyl,morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl,phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl,prolinyl, pteridinyl, purynyl, pyranyl, pyrazinyl, pyroazolidinyl,pyrazolinyl, pyrazolyl, pyridazinyl, pyridonyl, pyridooxazoles,pyridoimidazoles, pyridothiazoles, pyridinyl, pyridyl, pyrimidinyl,pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydrofuranyl,tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadazinyl,thiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl,1,3,4-thiadiazolyl, thienyl, triazolyl, tetrazolyl and xanthenyl.Pyridyl stands both for 2-, 3- and 4-pyridyl. Thienyl stands both for 2-and 3-thienyl. Furyl stands both for 2- and 3-furyl. Also included arethe corresponding N-oxides of these compounds, for example, 1-oxy-2-, 3-or 4-pyridyl.

Substitutions in (C₅-C₁₀)heterocyclyl residues can occur on free carbonatoms or on nitrogen atoms.

Preferred examples of (C₅-C₁₀)heterocyclyl residues are pyrazinyl,pyridyl, pyrimidinyl, pyrazolyl, morpholinyl, pyrrolidinyl, piperazinyl,piperidinyl, thienyl, benzofuryl, quinolinyl, tetrazolyl and triazolyl.

(C₆-C₁₀)aryl and (C₅-C₁₀)heterocyclyl groups are unsubstituted orsubstituted one or more times by suitable groups independently selectedfrom halogen, CF₃, NO₂, N₃, CN, C(O)—(C₁-C₆)alkyl, C(O)—(C₁-C₆)aryl,COOH, COO(C₁-C₆)alkyl, CONH₂, CONH(C₁-C₆)alkyl, CON[(C₁-C₆)alkyl]₂,(C₃-C₈)cycloalkyl, (C₁-C₆)alkyl, (C₁-C₆)alkylene-OH,(C₁-C₆)alkylene-NH₂, (C₁-C₆)alkylene-NH(C₁-C₆)alkyl,(C₁-C₆)alkylene-N[(C₁-C₆)alkyl]₂, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl,O—(C₁-C₆)alkyl, O—C(O)—(C₁-C₆)alkyl, O—C(O)—(C₆-C₁₀)aryl,O—C(O)—(C₅-C₁₀)heterocyclyl, PO₃H₂, SO₃H, SO₂—NH₂, SO₂NH(C₁-C₆)alkyl,SO₂N[(C₁-C₆)alkyl]₂, S—(C₁-C₆)alkyl; S—(C₁-C₆)alkylene-(C₆-C₁₀)aryl,S—(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl, SO—(C₁-C₆)alkyl,SO—(C₁-C₆)alkylene-(C₆-C₁₀)aryl,SO—(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl, SO₂—(C₁-C₆)alkyl,SO₂—(C₁-C₆)alkylene-(C₆-C₁₀)aryl,SO₂—(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl,SO₂—NH(C₁-C₆)alkylene-(C₆-C₁₀)aryl,SO₂—NH(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl,SO₂—N[(C₁-C₆)alkyl][(C₁-C₆)alkylene-(C₆-C₁₀)aryl],SO₂—N[(C₁-C₆)alkyl][(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl],SO₂—N[(C₁-C₆)alkylene-(C₆-C₁₀)aryl]₂,SO₂—N[(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl]₂,

C(NH)(NH₂), NH₂, NH—(C₁-C₆)alkyl, N[(C₁-C₆)alkyl]₂,NH—C(O)—(C₁-C₆)alkyl, NH—C(O)O—(C₁-C₆)alkyl, NH—C(O)—(C₆-C₁₀)aryl,NH—C(O)—(C₅-C₁₀)heterocyclyl, NH—C(O)O—(C₆-C₁₀)aryl,NH—C(O)O—(C₅-C₁₀)heterocyclyl, NH—C(O)—NH—(C₁-C₆)alkyl,NH—C(O)—NH—(C₆-C₁₀)aryl, NH—C(O)—NH—(C₅-C₁₀)heterocyclyl,NH—SO₂—(C₁-C₆)alkyl, NH—SO₂—(C₆-C₁₀)aryl, NH—SO₂—(C₅-C₁₀)heterocyclyl,N(C₁-C₆)alkyl-C(O)—(C₁-C₆)alkyl, N(C₁-C₆)alkyl-C(O)O—(C₁-C₆)alkyl,N(C₁-C₆)alkyl-C(O)—(C₆-C₁₀)aryl, N(C₁-C₆)alkyl-C(O)-heterocyclyl,N(C₁-C₆)alkyl-C(O)O—(C₆-C₁₀)aryl, N(C₁-C₆)alkyl-C(O)O—(C₅-C₁₀)heterocyclyl, N(C₁-C₆)alkyl-C(O)—NH—(C₁-C₆)alkyl],N(C₁-C₆)alkyl-C(O)—NH—(C₆-C₁₀)aryl,N(C₁-C₆)alkyl-C(O)—NH—(C₅-C₁₀)heterocyclyl,N[(C₁-C₆)alkyl]-C(O)—N[(C₁-C₆)alkyl]₂,N[(C₁-C₆)alkyl]-C(O)—N[(C₁-C₆)alkyl]-(C₆-C₁₀)aryl,N[(C₁-C₆)alkyl]-C(O)—N[(C₁-C₆)alkyl]-(C₅-C₁₀)heterocyclyl,N[(C₁-C₆)alkyl]-C(O)—N[(C₆-C₁₀)aryl]₂,N[(C₁-C₆)alkyl]-C(O)—N[(C₅-C₁₀)heterocyclyl]₂,N[(C₆-C₁₀)aryl]-C(O)—(C₁-C₆)alkyl,N[(C₅-C₁₀)heterocyclyl]-C(O)—(C₁-C₆)alkyl,N[(C₆-C₁₀)aryl]-C(O)O—(C₁-C₆)alkyl,N[(C₅-C₁₀)heterocyclyl]-C(O)O—(C₁-C₆)alkyl, N(aryl)-C(O)—(C₆-C₁₀)aryl,N[(C₅-C₁₀)heterocyclyl]-C(O)—(C₆-C₁₀)aryl,N[(C₆-C₁₀)aryl]-C(O)O—(C₆-C₁₀)aryl,N[(C₅-C₁₀)heterocyclyl]-C(O)O—(C₆-C₁₀)aryl,N[(C₆-C₁₀)aryl]-C(O)—NH—(C₁-C₆)alkyl,N[(C₅-C₁₀)heterocyclyl]-C(O)—NH—(C₁-C₆)alkyl,N(aryl)-C(O)—NH—(C₆-C₁₀)aryl,N[(C₅-C₁₀)heterocyclyl]-C(O)—NH—(C₆-C₁₀)aryl,N[(C₆-C₁₀)aryl]-C(O)—N[(C₁-C₆)alkyl]₂,N[(C₅-C₁₀)heterocyclyl]-C(O)—N[(C₁-C₆)alkyl]₂,N[(C₆-C₁₀)aryl]-C(O)—N[(C₁-C₆)alkyl]-(C₆-C₁₀)aryl,N[(C₅-C₁₀)heterocyclyl]-C(O)—N[(C₁-C₆)alkyl]-(C₆-C₁₀)aryl,N[(C₆-C₁₀)aryl]-C(O)—N[(C₆-C₁₀)aryl]₂,N[(C₅-C₁₀)heterocyclyl]-C(O)—N[(C₆-C₁₀)aryl]₂, (C₆-C₁₀)aryl,(C₁-C₆)alkylene-(C₆-C₁₀)aryl, O—(C₁-C₆)alkylene-(C₆-C₁₀)aryl,(C₅-C₁₀)heterocyclyl, (C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl,O—(C₁-C₆)alkylene-(C₅-C₁₀)heterocyclyl, wherein the (C₆-C₁₀)aryl or(C₅-C₁₀)heterocyclyl may be substituted one to 3 times by halogen, OH,NO₂, CN, O—(C₁-C₆)alkyl, (C₁-C₆)alkyl, NH₂, NH(C₁-C₆)alkyl,N[(C₁-C₆)alkyl]₂, SO₂CH₃, COOH, C(O)O—(C₁-C₆)alkyl, CONH₂,(C₁-C₆)alkylene-O—(C₁-C₆)alkyl, (C₁-C₆)alkylene-O—(C₆-C₁₀)aryl,O—(C₁-C₆)alkylene-(C₆-C₁₀)aryl; or wherein (C₆-C₁₀)aryl is vicinalsubstituted by a O—(C₁-C₄)alkylene-O group whereby a 5-8-membered ringis formed together with the carbon atoms the oxygen atoms are attachedto. Aryl or heterocyclyl substituents of (C₆-C₁₀)aryl and(C₅-C₁₀)heterocyclyl groups may not be further substituted by an aryl orheterocyclyl containing group.

Preferred substituents for (C₆-C₁₀)aryl groups are (C₁-C₄)alkyl,O—(C₁-C₄)alkyl, O-phenyl, C(O)O—(C₁-C₆)alkyl, C(O)OH, C(O)—(C₁-C₄)alkyl,halogen, NO₂, SO₂NH₂, CN, SO₂—(C₁-C₄)alkyl, NH—SO₂—(C₁-C₄)alkyl, NH₂,NH—C(O)—(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, (C₁-C₄)alkyl-OH,C(O)N[(C₁-C₄)alkyl]₂, C(O)NH₂, N[(C₁-C₄)alkyl]₂,(C₂-C₄)alkenylene-(C₆-C₁₀)aryl, wherein the (C₆-C₁₀)aryl may be furthersubstituted by (C₁-C₄)alkyl, (C₁-C₄)alkylene-O—(C₁-C₆)alkyl,O—(C₁-C₆)alkyl-(C₆-C₁₀)aryl, or may be vicinal substituted by aO—(C₁-C₄)alkylene-O group whereby a 5-8-membered ring is formed togetherwith the carbon atoms the oxygen atoms are attached to.

In monosubstituted phenyl groups the substituent can be located in the2-position, the 3-position or the 4-position, with the 3-position andthe 4-position being preferred. If a phenyl group carries twosubstituents, they can be located in 2,3-position, 2,4-position,2,5-position, 2,6-position, 3,4-position or 3,5-position. In phenylgroups carrying three substituents the substituents can be located in2,3,4-position, 2,3,5-position, 2,3,6-position, 2,4,5-position,2,4,6-position, or 3,4,5-position.

The above statements relating to phenyl groups correspondingly apply todivalent groups derived from phenyl groups, i.e. phenylene which can beunsubstituted or substituted 1,2-phenylene, 1,3-phenylene or1,4-phenylene. The above statements also correspondingly apply to thearyl subgroup in arylalkylene groups. Examples of arylalkylene groupswhich can also be unsubstituted or substituted in the aryl subgroup aswell as in the alkylene subgroup, are benzyl, 1-phenylethylene,2-phenylethylene, 3-phenylpropylene, 4-phenylbutylene,1-methyl-3-phenyl-propylene.

Preferred substituents for (C₅-C₁₀)heterocyclyl groups are (C₁-C₄)alkyl,O—(C₁-C₄)alkyl, (C₁-C₄)alkylene-phenyl, halogen,(C₁-C₄)alkylene-O—(C₁-C₄)alkyl, (C₅-C₁₀)heterocyclyl,(C₁-C₄)alkylene-N[(C₁-C₄)alkyl]₂, or (C₆-C₁₀)aryl, wherein the(C₆-C₁₀)aryl may be further substituted by (C₁-C₄)alkyl,(C₁-C₄)alkylene-O—(C₁-C₆)alkyl, O—(C₁-C₆)alkyl-(C₆-C₁₀)aryl, or may bevicinal substituted by a O—(C₁-C₄)alkylene-O group whereby a5-8-membered ring is formed together with the carbon atoms the oxygenatoms are attached to.

The general and preferred substituents of (C₆-C₁₀)aryl and(C₅-C₁₀)heterocyclyl groups may be combined with the general andpreferred definitions of R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈, n and L asdescribed above.

The present invention therefore also relates to the compounds of theformula (I) and/or their physiologically acceptable salts and/or theirprodrugs for use as pharmaceuticals (or medicaments), to the use of thecompounds of the formula (I) and/or their physiologically acceptablesalts and/or their prodrugs for the production of pharmaceuticals forthe treatment and/or prevention of diseases associated with Rho-kinaseand/or Rho-kinase mediated phosphorylation of myosin light chainphosphatase, i.e. for the treatment and/or prevention of hypertension,i.e. pulmonary hypertension and ocular hypertension, peripheralcirculatory disorder, angina pectoris, cerebral vasospasm, asthma,premature birth, hyperaggregability of platelets, Peripheral OcclusiveArterial Disease (PAOD), Chronic Obstructive Pulmonary Disease (COPD),cancer development, erectile dysfunction, arteriosclerosis, ischemicorgan failure (end organ damage), fibroid lung, fibroid liver, liverfailure, fibroid kidney, renal glomerulosclerosis, kidney failure, organhypertrophy, prostatic hypertrophy, complications of diabetes, bloodvessel restenosis, atherosclerosis, cancer, cardiac hypertrophy, heartfailure; ischemic diseases; inflammation; autoimmune diseases; AIDS,osteopathy such as osteoporosis, brain functional disorder, infection ofdigestive tracts with bacteria, sepsis, adult respiratory distresssyndrome, retinopathy, glaucoma and Alzheimer's disease.

The present invention furthermore relates to pharmaceutical preparations(or pharmaceutical compositions) which contain an effective amount of atleast one compound of the formula (I) and/or its physiologicallyacceptable salts and/or its prodrugs and a pharmaceutically acceptablecarrier, i.e. one or more pharmaceutically acceptable carrier substances(or vehicles) and/or additives (or excipients).

The pharmaceuticals can be administered orally, for example in the formof pills, tablets, lacquered tablets, coated tablets, granules, hard andsoft gelatin capsules, solutions, syrups, emulsions, suspensions oraerosol mixtures. Administration, however, can also be carried outrectally, for example in the form of suppositories, or parenterally, forexample intravenously, intramuscularly or subcutaneously, in the form ofinjection solutions or infusion solutions, microcapsules, implants orrods, or percutaneously or topically, for example in the form ofointments, solutions or tinctures, or in other ways, for example in theform of aerosols or nasal sprays.

The pharmaceutical preparations according to the invention are preparedin a manner known per se and familiar to one skilled in the art,pharmaceutically acceptable inert inorganic and/or organic carriersubstances and/or additives being used in addition to the compound(s) ofthe formula (I) and/or its (their) physiologically acceptable saltsand/or its (their) prodrugs. For the production of pills, tablets,coated tablets and hard gelatin capsules it is possible to use, forexample, lactose, corn starch or derivatives thereof, talc, stearic acidor its salts, etc. Carrier substances for soft gelatin capsules andsuppositories are, for example, fats, waxes, semisolid and liquidpolyols, natural or hardened oils, etc. Suitable carrier substances forthe production of solutions, for example injection solutions, or ofemulsions or syrups are, for example, water, saline, alcohols, glycerol,polyols, sucrose, invert sugar, glucose, vegetable oils, etc. Suitablecarrier substances for microcapsules, implants or rods are, for example,copolymers of glycolic acid and lactic acid. The pharmaceuticalpreparations normally contain about 0.5 to about 90% by weight of thecompounds of the formula (I) and/or their physiologically acceptablesalts and/or their prodrugs. The amount of the active ingredient of theformula (I) and/or its physiologically acceptable salts and/or itsprodrugs in the pharmaceutical preparations normally is from about 0.5to about 1000 mg, preferably from about 1 to about 500 mg.

In addition to the active ingredients of the formula (I) and/or theirphysiologically acceptable salts and/or prodrugs and to carriersubstances, the pharmaceutical preparations can contain one or moreadditives such as, for example, fillers, disintegrants, binders,lubricants, wetting agents, stabilizers, emulsifiers, preservatives,sweeteners, colorants, flavorings, aromatizers, thickeners, diluents,buffer substances, solvents, solubilizers, agents for achieving a depoteffect, salts for altering the osmotic pressure, coating agents orantioxidants. They can also contain two or more compounds of the formula(I) and/or their physiologically acceptable salts and/or their prodrugs.In case a pharmaceutical preparation contains two or more compounds ofthe formula (I) the selection of the individual compounds can aim at aspecific overall pharmacological profile of the pharmaceuticalpreparation. For example, a highly potent compound with a shorterduration of action may be combined with a long-acting compound of lowerpotency. The flexibility permitted with respect to the choice ofsubstituents in the compounds of the formula (I) allows a great deal ofcontrol over the biological and physico-chemical properties of thecompounds and thus allows the selection of such desired compounds.Furthermore, in addition to at least one compound of the formula (I)and/or its physiologically acceptable salts and/or its prodrugs, thepharmaceutical preparations can also contain one or more othertherapeutically or prophylactically active ingredients.

When using the compounds of the formula (I) the dose can vary withinwide limits and, as is customary and is known to the physician, is to besuited to the individual conditions in each individual case. It depends,for example, on the specific compound employed, on the nature andseverity of the disease to be treated, on the mode and the schedule ofadministration, or on whether an acute or chronic condition is treatedor whether prophylaxis is carried out. An appropriate dosage can beestablished using clinical approaches well known in the medical art. Ingeneral, the daily dose for achieving the desired results in an adultweighing about 75 kg is from about 0.01 to about 100 mg/kg, preferablyfrom about 0.1 to about 50 mg/kg, in particular from about 0.1 to about10 mg/kg, (in each case in mg per kg of body weight). The daily dose canbe divided, in particular in the case of the administration ofrelatively large amounts, into several, for example 2, 3 or 4, partadministrations. As usual, depending on individual behavior it may benecessary to deviate upwards or downwards from the daily dose indicated.

Furthermore, the compounds of the formula (I) can be used as synthesisintermediates for the preparation of other compounds, in particular ofother pharmaceutical active ingredients, which are obtainable from thecompounds of the formula I, for example by introduction of substituentsor modification of functional groups.

The compounds of the formula (I) can be prepared according to thefollowing exemplified compounds without limiting the scope of theclaims.

In general, protective groups that may still be present in the productsobtained in the coupling reaction are then removed by standardprocedures. For example, tert-butyl protecting groups, in particular atert-butoxycarbonyl group which is a protected form of an amidino group,can be deprotected, i.e. converted into the amidino group, by treatmentwith trifluoroacetic acid. As already explained, after the couplingreaction also functional groups can be generated from suitable precursorgroups. In addition, a conversion into a physiologically acceptable saltor a prodrug of a compound of the formula (I) can then be carried out byknown processes.

In general, a reaction mixture containing a final compound of theformula (I) or an intermediate is worked up and, if desired, the productis then purified by customary processes known to those skilled in theart. For example, a synthesized compound can be purified using wellknown methods such as crystallization, chromatography or reversephase-high performance liquid chromatography (RP-HPLC) or other methodsof separation based, for example, on the size, charge or hydrophobicityof the compound. Similarly, well known methods such as amino acidsequence analysis, NMR, IR and mass spectrometry (MS) can be used forcharacterizing a compound of the invention.

It is understood that modifications that do not substantially affect theactivity of the various embodiments of this invention are includedwithin the invention disclosed herein. Accordingly, the followingexamples are intended to illustrate but not limit the present invention.

LCMS Methods

Method #1

Column: YMC J'shere 33×2 4 μm

gradient (AcN+0.05% TFA): H2O+0.05% TFA; 5:95 (0 min) to 95:5 (2.5 min)to 95:5 (3 min)

Method #2

Column: YMC J'shere 33×2 4 μm

gradient (AcN+0.05% TFA): H2O+0.05% TFA, 5:95 (0 min) to 95:5 (3.4 min)to 95:5 (4.4 min)

Method #3

Column: YMC J'shere 33×2 4 μm

gradient AcN+0.08% FA: H2O+0.1% FA; 5:95 (0 min) to 95:5(2.5 min) to95:5(3 min)

Method #Top

Column: YMC YMC J'sphere ODS H80 20×2 1 4μ

gradient 0 min 96% H2O (0.05% TFA) 2.0 min-95% ACN; 95% ACN bis 2.4 min;4% ACN 2.45 min

Building Block Syntheses

7-Bromo-isoquinoline-6-ol (1)

25 g (116.3 mmol) of 3-bromo-4-methoxybenzaldehyde, 19.0 mL (18.3 g,174.5 mmol) of aminoacetaldehyde dimethyl acetal and 250 mL of toluenewere heated to reflux for 6 h using a Dean-Stark apparatus. Solvent andexcess reagent were distilled off and the crude product (approx. 37 g)was used for the next step without any additional purification.

The imine was dissolved in 240 mL of THF. 11.1 mL (12.6 g, 116.3 mmol)of ethyl chloroformate were added dropwise at 0° C. After stirring for 5minutes 24.3 mL (23.2 g, 139.2 mmol) triethylphosphite were addeddropwise. The mixture was stirred for 18 h at room temperature. Then thesolvents were distilled off. Excess reagent was removed by repeatedaddition of 100 ml toluene and evaporation of the solvents. TheP,N-acetal (approx. 62 g) was used for the next step without anyadditional purification.

The P,N-acetal, 51.3 mL (88.2 g, 465.2 mmol) titanium tetrachloride and300 mL chloroform were heated to reflux for 48 h. The mixture was pouredon ice and the pH was adjusted to 9 by using aqueous ammonia. Repeatedextraction with ethyl acetate followed by removal of the solvents gave14.8 g (53%) of 7-bromo-6-methoxyisoquinoline.

¹H-NMR (d₆-DMSO): δ=9.16 (1H, s), 8.46 (1H, d, J=5.9 Hz), 8.46 (1H, s),7.76 (1H, d, J=5.9 Hz), 7.51 (1H, s), 4.01 (3H, s).

MS: m/z=238 (MH⁺).

3.6 mL (9.5 g, 37.8 mmol) of BBr₃ were added at 0° C. to a solution of4.5 g (18.9 mmol) 7-bromo-6-methoxy isoquinoline in 30 mLdichloromethane and stirred for 18 h at room temperature. AqueousNaHCO₃-solution was added to adjust the pH to 8. Extraction withchloroform/isopropanol (3/1) followed by drying over sodium sulfate andremoval of the solvents gave 2.7 g (64%) of compound 1.

¹H-NMR (d₆-DMSO): δ=9.19 (1H, s), 8.49 (1H, s), 8.38 (1H, d, J=6.1 Hz),7.78 (1H, d, J=6.1 Hz), 7.34 (1H, s).

MS: m/z=224 (MH⁺).

The following intermediates were synthesized using this procedure:

8-Fluoro-isoquinoline-6-ol (2)

¹H-NMR (d₆-DMSO): δ=10.84 (1H, s), 9.21 (1H, s), 8.40 (1H, d, J=5.8 Hz),7.67 (1H, d, J=5.8 Hz), 7.01 (2H, m).

MS: m/z=164 (MH⁺).

7-Fluoro-isoquinoline-6-ol (3)

¹H-NMR (d₆-DMSO): δ=11.06 (1H, s), 9.07 (1H, s), 8.33 (1H, d, J=5.6 Hz),7.88 (1H, d, J=11.4 Hz), 7.64 (1H, d, J=5.6 Hz), 7.31 (1H, d, J=8.6 Hz).

MS: m/z=164 (MH⁺).

8-Methyl-isoquinoline-6-ol (4)

¹H-NMR (d₆-DMSO): δ=11.55 (1H, s), 9.47 (1H, s), 8.42 (1H, d, J=6.5 Hz),8.11 (1H, d, J=6.5 Hz), 7.31 (1H, s), 7.25 (1H, s), 2.76 (3H, s).

MS: m/z=160 (MH⁺).

7,8-Dimethyl-isoquinoline-6-ol (5)

¹H-NMR (d₆-DMSO): δ=11.87 (1H, s), 9.58 (1H, s), 8.41 (1H, d, J=6.5 Hz),8.18 (1H, d, J=6.5 Hz), 7.35 (1H, s), 7.25 (1H, s), 2.71 (3H, s), 2.35(3H, s).

MS: m/z=174 (MH⁺).

5,8-Dimethyl-isoquinoline-6-ol (6)

¹H-NMR (d₆-DMSO): δ=11.55 (1H, s), 9.52 (1H, s), 8.47 (1H, d, J=6.8 Hz),8.26 (1H, d, J=6.8 Hz), 7.42 (1H, s), 2.76 (3H, s), 2.42 (3H, s).

MS: m/z=174 (MH⁺).

6-Hydroxy-isoquinoline (7)

LCMS Method #1, retention time 0.14 min, detected mass 146.08 [M+H]⁺

5-Chloroisoquinoline-6-ol (8)

0.61 mL (1.02 g, 7.6 mmol) of sulfuryl chloride were added to a solutionof 1.0 g (6.9 mmol) of compound 7 in 30 mL of dichloromethane. Threedrops diethyl ether were added and the reaction was stirred at roomtemperature for 5 h. The solvents were removed by distillation and theremainder was treated with aqueous NaHCO₃ solution. The precipitate wasfiltered, washed with water and dried to give 1.1 g (89%) of compound 8as a green-yellow solid.

¹H-NMR (d₆-DMSO): δ=11.37 (1H, s), 9.18 (1H, s), 8.50 (1H, d, J=6 Hz),8.00 (1H, d, J=8.8 Hz), 7.83 (1H, J=6 Hz), 7.44 (1H, d, J=8.7 Hz).

MS: m/z=180 (MH⁺).

5-Bromoisoquinoline-6-ol (9)

7.9 mL (19.18 g, 120 mmol) of bromine were added dropwise to asuspension of 17.42 g (120 mmol) of compound 7 in 250 mL of chloroformat room temperature. After stirring for 2 h ethyl acetate was added. Theprecipitate was filtered, washed with ethyl acetate and dried. AqueousNaHCO₃ solution was added carefully. The precipitate was filtered andwashed with NaHCO₃ solution until the filtrate had a pH of 8. Dryinggave 23.78 g (88%) of compound 9 as an off-white solid.

¹H-NMR (d₆-DMSO): δ=11.30 (1H, s), 9.13 (1H, s), 8.48 (1H, d, J=5.9 Hz),8.02 (1H, d, J=8.8 Hz), 7.78 (1H, J=5.9 Hz), 7.40 (1H, d, J=8.8 Hz).

MS: m/z=224 (MH⁺).

5-Iodoisoquinoline-6-ol (10)

Under argon atmosphere 1.77 g (12.2 mmol) of compound 7 were added to asolution of 5.0 g (13.5 mmol) bis(pyridin)iodonium tetrafluoroborate in100 mL of dry dichloromethane. A solution of 2.4 mL (4 g, 26.8 mmol)trifluoromethane sulfonic acid in 20 mL dry dichloromethane was addeddropwise at 0° C. and the mixture was stirred for 3 hours at roomtemperature. The solvents were removed by distillation and the remainderwas treated with aqueous NaHCO₃ solution. The precipitate was filtered,washed with water and dried to yield 3.2 g (97%) of compound 10 as abeige solid.

¹H-NMR (d₆-DMSO): δ=9.09 (1H, s), 8.47 (1H, d, J=6.1 Hz), 8.04 (1H, d,J=8.8 Hz), 7.76 (1H, J=6.1 Hz), 7.37 (1H, d, J=8.8 Hz).

MS: m/z=272 (MH⁺).

4-(5-Bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (11)

3.75 mL (4.15 g, 23.8 mmol) of diethyl azo dicarboxylate were added to12.7 g (19.9 mmol) of polymer-bound triphenylphosphine (PS—PPh₃, approx.1.6 mmol/g, Argonaut) in 250 mL of dichloromethane at 0° C. and stirredfor 15 min. 4.45 g (19.9 mmol) 5-bromo isoquinoline-6-ol (9), 4.0 g(19.9 mmol) Boc-(4-hydroxy)piperidine and 4.1 mL (3.0 g, 29.8 mmol)triethyl amine were added. The mixture was shaken for 16 h. The polymerwas removed by filtration through Celite and the solvents were distilledoff. 20 mL dichloromethane were added and the precipitate was isolatedby filtration. The crude product (8 g) was purified by flashchromatography using ethyl acetate/n-heptane as eluent to give 4.78 g(60%) of compound 11.

¹H-NMR (d₆-DMSO): δ=9.24 (1H, s), 8.97 (1H, s), 8.56 (1H, d, J=6 Hz),8.20 (1H, d, J=9 Hz), 7.85 (1H, d, J=6 Hz), 7.75 (1H, d, J=9 Hz), 5.02(1H, m), 3.58 (2H, m), 3.40 (2H, m), 1.91 (2H, m), 1.70 (2H, m), 1.41(9H, s).

MS: m/z=407 (MH⁺).

The following building blocks were synthesized according to this method:

4-(5-Iodo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (12)

using compound 10 as starting material

¹H-NMR (CDCl₃): δ=9.04 (1H, s), 8.55 (1H, d, J=6 Hz), 7.93 (1H, d, J=9Hz), 7.86 (1H, d, J=6 Hz), 7.27 (1H, d, J=9 Hz), 4.87 (1H, m), 3.66 (4H,m), 1.93 (4H, m), 1.48 (9H, s).

MS: m/z=455 (MH⁺).

4-(7-Bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (13)

using compound 1 as starting material

LCMS Method #4, retention time 1.13 min, detected mass 407.4 [M+H]⁺

4-[5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-isoquinoline-6-yloxy]-piperidin-1-carboxylicacid tert-butyl ester (14)

A solution of 0.55 g (1.34 mmol)4-(5-bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (11) in 14 mL of DMSO was added to a mixture of 1.0 g (4.0 mmol)bis(pinacolato)diboron, 0.78 g (8.0 mmol) K₂CO₃ and 29 mg (0.03 eq.)Pd(dppf)Cl₂. Argon was bubbled through the mixture for 30 min and thenthe reaction mixture was heated in a microwave reactor (CEM Discovery)to 100° C. for 60 min. After cooling to room temperature water wasadded. The mixture was extracted with ethyl acetate. After removal ofthe solvent the product was isolated by flash chromatography (ethylacetate/n-heptane) to yield: 269 mg (44%) of compound 14 as a whitesolid. LCMS Method #4, retention time 1.30 min, detected mass 433.3[M+H]⁺

4-(5-Cyano-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (15)

Under argon atmosphere 47 mg (0.4 mmol) of Zn(CN)₂ and 23 mg of (0.02eq) Pd(PPh₃)₄ were added to a solution 62 mg (0.4 mmol) of4-(5-bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (11) in DMF. The reaction was heated for 5 minutes to 150° C. in amicrowave reactor (CEM Discovery). After cooling to room temperaturewater and ethyl acetate were added. The mixture was filtered throughcelite, washed with ethyl acetate and concentrated to yield 176 mg ofcompound 15.

MS: m/z=354 (MH⁺).

4-(7-Cyano-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (16)

Under argon atmosphere 35 mg (0.3 mmol) of Zn(CN)₂ and 17 mg (0.05 eq)of Pd(PPh₃)₄ were added to a solution of 122 mg (0.3 mmol) of4-(7-bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (13) in DMF. The reaction was heated for 5 minutes to 150° C. in amicrowave reactor (CEM Discovery). After cooling to room temperaturewater and ethyl acetate were added. The mixture was filtered throughCelite, washed with ethyl acetate and concentrated. The crude productwas purified by preparative HPLC to yield 77 mg of compound 16.

LCMS Method #4, retention time 1.06 min, detected mass 354.5 [M+H]⁺

4-(5-Azido-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (17)

Under argon atmosphere 40 μL (0.04 mmol) of 1N NaOH, 4.6 mg (0.04 mmol)of L-proline, 3.8 mg of (0.02 mmol) CuI and 15.6 mg (0.24 mmol) of NaN₃were added to a solution of 91 mg (0.2 mmol) of4-(5-iodo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (12) in 2 mL of DMSO. The mixture was heated to 60° C. for 18 h.NaN₃, NaOH and L-proline were added in the same amounts again and thereaction was heated to 60° C. for 5 h. After cooling to room temperaturewater was added. The precipitate was filtered, washed with water anddried in vacuo to give 74 mg of compound 17, which was used without anyadditional purification.

MS: m/z=370 (MH⁺).

4-(5-Amino-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (18)

Under argon atmosphere 600 μL (0.6 mmol) of 1N NaOH, 13.8 mg (0.12 mmol)L-proline, 7.6 mg (0.04 mmol) of CuI and 52 mg (0.8 mmol) of NaN₃ wereadded to a solution of 163 mg (0.4 mmol)4-(5-bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11) in 0.6 mL of water. The mixture was heated to 95° C. for 3 hin a microwave reactor (CEM Discovery). After cooling to roomtemperature water and ethyl acetate were added. The mixture was filteredthrough Celite, washed with ethyl acetate and concentrated. The crudeproduct was purified by preparative HPLC to yield 42 mg of compound 18(containing some 11 as impurity).

LCMS Method #4, retention time 0.97 min, detected mass 344.5 [M+H]⁺

4-(7-Vinyl-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (19)

Under argon atmosphere 340 mg tributyl-vinyl-stannane (1.07 mmol, 1.2eq.) and 103 mg of Pd(PPh₃)₄ (0.1 eq.) were added to a solution of 364mg of 4-(7-Bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acidtert-butyl ester (13) (0.98 mmol) in 4 ml of toluene. The reaction washeated to 100° C. in a microwave reactor (CEM Discovery) for 1 h.

After cooling to room temperature water and ethyl acetated were added.The mixture was filtered through a Celite cartridge, washed with ethylacetate and concentrated. The crude product was purified by preparativeHPLC to yield 256 mg (81%) of compound 19.

LCMS Method #4, retention time 1.19 min, detected mass 355.5 [M+H]⁺

The following building blocks were synthesized according to this method:

4-(5-Vinyl-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (19A)

using compound 11 as starting material

LCMS Method #4, retention time 1.11 min, detected mass 355.4 [M+H]⁺

4-(7-Thiophen-2-yl-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (20)

using compound 13 and tributyl-thiophen-2-yl-stannane as startingmaterials.

LCMS Method #4, retention time 1.26 min, detected mass 411.5 [M+H]⁺

(2,2-Dimethoxy-ethyl)-(4-fluoro-benzyl)-amine (21)

12.4 g of 4-Fluorobenzaldehyde were dissolved in 100 mL of toluene andreacted with 10.5 g 2-Aminoacetaldehyde dimethylacetal and 1.90 g (10mmol) p-toluenesulfonic acid monohydrate for two hours at a Dean Starkapparatus. The solution was allowed to cool down, extracted withsaturated sodium bicarbonate, water and brine, dried over magnesiumsulfate and evaporated to dryness. The crude product was dissolved in100 mL of ethanol. 1.89 g of sodium borohydride were added portionwise.Stirring was continued overnight. For workup, acetic acid was addeduntil no gas evolution could be observed. Then the solution wasevaporated to dryness, taken up in dichloromethane and washed twice withwater. The organic layer was extracted with brine, dried over magnesiumsulfate and evaporated to dryness. The obtained crude product (20 g) wasused for further reactions without purification. R_(t)=0.86 min (Method#1). Detected mass: 182.1 (M-OMe⁻), 214.2 (M+H⁺).

N-(2,2-Dimethoxy-ethyl)-N-(4-fluoro-benzyl)-4-methyl-benzenesulfonamide(22)

20 g (2,2-Dimethoxy-ethyl)-(4-fluoro-benzyl)-amine (21) were dissolvedin 120 ml of dichloromethane. 20 mL of pyridine are added. At 0° C. asolution of 23.8 g p-toluenesulfonic acid chloride in dichloromethanewas added dropwise. The reaction was allowed to warm to room temperatureand stirring is continued until conversion was completed. For workup,the reaction mixture was extracted twice with 2M hydrochloric acid,twice with sodium bicarbonate and once with brine. The organic layer wasdried over magnesium sulfate, evaporated to dryness and the obtainedcrude product was purified by silica gel chromatography to yield 22.95 gof compound 22 as an orange oil. R_(t)=1.71 min (Method #4). Detectedmass: 336.1 (M-OMe⁻).

6-Fluoro-isoquinoline (23)

41.6 g of AlCl₃ were suspended in 400 mL of dichloromethane. At roomtemperature, a solution of 22.95 g ofN-(2,2-Dimethoxy-ethyl)-N-(4-fluoro-benzyl)-4-methyl-benzenesulfonamide(22) in 150 ml of dichloromethane was added. Stirring was continued atroom temperature overnight, the solution was poured on ice, the organiclayer was separated, the aqueous phase was extracted twice withdichloromethane and the combined organic layers are then extracted twicewith sodium bicarbonate. The organic layer was dried over magnesiumsulfate, evaporated to dryness and the obtained crude product (8.75 g)is purified by silica gel chromatography to yield 2.74 g of compound 23.R_(t)=0.30 min (Method #4). Detected mass: 148.1 (M+H⁺).

4-Chloro-6-fluoro-isoquinoline (24)

A solution of 1.5 g 6-fluoro-isoquinoline (23) in 4.5 ml sulfurylchloride was heated to 60° C. in a microwave reactor (CEM Discovery) for8 h. After cooling to room temperature the mixture was poured on ice andextracted three times with CHCl₃. After drying over Na₂SO₄ the solventwas distilled off and the crude product was purified by flashchromatography to yield 930 mg of compound 24.

LCMS Method #1, retention time 1.37 min, detected mass 182.01 [M+H]⁺

Cis and trans N-Boc-2-methyl-piperidin-4-ol (25 and 26)

213 mg (5.6 mmol) of NaBH₄ were added portionwise at 0° C. to a solutionof 1.0 g (4.7 mmol) 1-Boc-2-methyl-piperidin-4-on in 10 mL EtOH. Themixture was stirred at room temperature for another 2 h. The solvent wasremoved by distillation and the remainder was dissolved in water andethyl acetate. The aqueous layer was extracted twice with ethyl acetatedand the combined organic layers were dried over Na₂SO₄. After filtrationthe solvent was removed by distillation and the crude product waspurified by column chromatography n-heptane/ethyl acetate (1/1) to yield367 mg (36%) of the cis-isomer 25 and 205 mg (20%) of the trans-isomer26 in addition to 97 mg (10%) mixture of both isomers.

Cis-Isomer (25):

¹H-NMR (CDCl₃): δ=4.28 (1H, m), 4.17 (1H, m), 3.82 (1H, m), 3.26 (1H,m), 1.85 (1H, ddd, J=14.7, 6.6, und 3.4 Hz), 1.77 (1H, m), 1.66 (2H, m),1.33 (3H, d, J=7.1 Hz).

Trans-Isomer (26):

¹H-NMR (CDCl₃): δ=4.50 (1H, m), 4.04 (1H, m), 3.95 (1H, m), 2.87 (1H,dt, J=2.9 und 13.6 Hz), 1.93 (1H, m), 1.83 (1H, m), 1.53 (1H, m), 1.32(1H, m), 1.14 (3H, d, J=7.1 Hz).

Phenyl-[6-(piperidin-4-yloxy)-isoquinolin-5-yl]-amine (27)

Under an argon atmosphere 81 mg (0.2 mmol) of4-(5-bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (11) and 24 mg (0.26 mmol) of aniline were added to a solution of27 mg (0.28 mmol) of NaOtBu in 3 mL of toluene. After stirring at roomtemperature for 10 min., 9 mg (0.05 eq) of Pd₂ dba₃ were added and themixture was heated to 100° C. in a microwave reactor (CEM Discovery) for1 h. After cooling to room temperature water and ethyl acetate wereadded. The organic layer was separated, dried over Na₂SO₄ andconcentrated. HPLC purification gave the Boc protected intermediatewhich was treated with 2 mL 5-6 N HCl in isopropanol for 2 h. Thehydrochloride was filtered and subjected to another HPLC chromatographyto yield compound 27 as trifluoroacetate (31.3 mg).

LCMS Method #2, retention time 0.78 min, detected mass 320.26 [M+H]⁺

5-Methyl-6-(piperidin-4-yloxy)-isoquinoline hydrochloride (28)

Under an argon atmosphere a 2 M solution of dimethyl zinc (0.5 mL, 93.7mg, 4 eq.) in toluene was added to a solution of 100 mg (0.24 mmol)4-(5-bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11) and 10 mg(1,1′-bis(diphenylphosphino)ferrocen)palladium(II)chloride (0.056 eq.Pd(dppf)Cl₂) in 3 mL of dioxane. The mixture was heated to 100° C. for 5h. After cooling the solvents were distilled off and the remainder wassubjected to preparative HPLC to give the Boc-protected intermediatewhich was treated with 5-6 N HCl in isopropanol for 2 h at roomtemperature. Removal of the solvents gave 13.7 mg (18%) of compound 28.

LCMS Method #1, retention time 0.67 min, detected mass 243.24 [M+H]⁺

5-Benzyl-6-(piperidin-4-yloxy)-isoquinoline (29)

0.3 mL of water were added to a solution of 81 mg (0.2 mmol) of4-(5-bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11), 195 mg (0.6 mmol) of Cs₂CO₃, 14.6 mg (0.02 mmol) ofPd(dppf)Cl₂ and 51 mg (0.26 mmol) of potassium benzyltrifluoroborate in3 mL of THF. Argon was bubbled through the mixture for 10 minutes andthen the reaction was heated to reflux for 16 h (incomplete conversion).After cooling to room temperature water and ethyl acetate were added.The organic layer was separated, dried over Na₂SO₄. After removal of thesolvents 2 mL 5-6 N HCl in isopropanol was added. After 2 h the solventswere distilled off and the remainder was subjected twice to preparativeHPLC to give 3.5 mg compound 29 as trifluoroacetate.

LCMS Method #3, retention time 0.56 min, detected mass 319.23 [M+H]⁺

6-(Piperidin-4-yloxy)-isoquinoline-5-carboxylic acid ethyl ester (30)

A solution of 200 mg (0.44 mmol)4-(5-iodo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (12), 107 mg (0.88 mmol) of DMAP, 4.7 mg (0.1 eq) of Pd oncharcoal (10%), 150 μL (0.88 mmol) of triethyl amine and 58 mg (0.22mmol) of Mo(CO)₆ in 3 mL of ethanol was heated to 135° C. for 1 h in amicrowave reactor (CEM Discovery). Then water and ethyl acetate wereadded and the mixture was filtered through a Celite cartridge. Afterremoval of the solvents the remainder was subjected to preparative HPLCto give the 7.4 mg Boc-protected intermediate. To remove the Boc groupthe intermediate was treated with 2 mL 5-6 N HCl in isopropanol at roomtemperature for 2 h. Purification by preparative HPLC gave 2.5 mg ofcompound 30 as TFA salt.

LCMS Method #3, retention time 0.14 min, detected mass 301.29 [M+H]⁺

6-(Piperidin-4-yloxy)-isoquinolin-5-ylamine (31)

60 μL of 1N NaOH-solution, 6.9 mg (0.3 eq) of L-proline, 3.8 mg (0.1 eq)of CuI and 26 mg (0.4 mmol) of NaN₃ were added to a solution of 82 mg(0.2 mmol) of 4-(5-Bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylicacid tert-butyl ester (11) in 2 mL of ethanol/water (7/3). The mixturewas heated to 95° C. for 3 h in a microwave reactor (CEM Discover).After cooling water and ethyl acetate were added and the mixture wasfiltered through a celite cartridge. After removal of the solvents bydistillation the remainder was subjected to preparative HPLC. TheN-Boc-protected intermediate was deprotected by treatment with 2 mL 5-6N HCl in isopropanol for 2 h at room temperature. Then water was addedand all solvents were removed by freeze drying to yield 18 mg ofcompound 31 as hydrochloride.

¹H-NMR (d₆-DMSO): δ=9.60 (1H, s), 8.95 (2H, br s), 8.56 (1H, d, J=7.1Hz), 8.41 (1H, d, J=7.1 Hz), 7.85 (1H, d, J=9.0 Hz), 7.81 (1H, d, J=9.0Hz), 5.03 (1H, m), 3.13 (1H, m), 2.92 (1H, m), 2.15 (2H, m), 1.99 (2H,m), 1.84 (1H, m), 1.55 (1H, m).

LCMS Method #1, retention time 0.35 min, detected mass 244.25 [M+H]⁺

6-(Piperidin-4-yloxy)-5-(1H-tetrazol-5-yl)-isoquinoline (32)

Under argon atmosphere 78 mg (1.2 mmol) of NaN₃ and 64 mg (1.2 mmol) ofNH₄Cl were added to a solution of 35 mg (0.1 mmol)4-(5-Cyano-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (15) in 1 mL of DMF. The mixture was heated to approximately 160°C. and 7 bar pressure for 3 h in a microwave reactor (CEM Discovery).After cooling to room temperature aqueous NH₄Cl-solution anddichloromethane was added. The mixture was filtered through a phaseseparation cartridge and the aqueous layer was washed twice withdichloromethane. The organic layers were combined and the solvents weredistilled off. The remainder was subjected to preparative HPLC to yield4 mg (8%) of compound 32 as trifluoroacetate. LCMS Method #3, retentiontime 0.90 min, detected mass 297.04 [M+H]⁺

5-(4-Methoxymethyl-[1,2,3]triazol-1-yl)-6-(piperidin-4-yloxy)-isoquinoline(33)

4 mg (0.1 eq.) of sodium ascorbate and 0.5 mg (0.01 eq.) ofCopper(II)sulfate-hydrate were added to a solution of 73 mg (0.2 mmol)of 4-(5-Azido-isoquinoline-6-yloxy)-piperidin-1-carboxylic acidtert-butyl ester (17) and 14 mg (0.2 mmol) of methyl propargyl ether in4 ml of water/tert-butanol (1/1). The mixture was stirred for 18 h atroom temperature. Then ethyl acetate was added and the mixture wasfiltered through a Celite cartridge. After removal of the solvents theremainder was subjected to preparative HPLC. The N-Boc-protectedintermediate was deprotected by treatment with 2 mL 5-6 N HCl inisopropanol for 2 h at room temperature. Then the solvent was evaporatedand the product was isolated by preparative HPLC to yield 2.8 mgcompound 33 as trifluoroacetate.

LCMS Method #3, retention time 0.08 min, detected mass 340.17 [M+H]⁺

5-(4-Phenyl-[1,2,3]triazol-1-yl)-6-(piperidin-4-yloxy)-isoquinoline (34)

According to the procedure described for compound 33 the title compoundwas obtained using 20 mg (0.2 mmol) phenylacetylene. Yield 2.5 mg ofcompound 34 as trifluoroacetate.

LCMS Method #3, retention time 0.14 min, detected mass 372.2 [M+H]⁺

7-Ethyl-6-(piperidin-4-yloxy)-isoquinoline hydrochloride (35)

1 mg of 5% Palladium on charcoal (0.02 eq.) was added to a solution of174 mg 4-(7-Vinyl-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (19) (0.49 mmol, 1 eq.) in 15 mL of methanol. Theolefin was hydrogenated under 5 bar H₂ at ambient temperature overnight. Only partial conversion was observed, thus the catalyst wasremoved by filtration and fresh catalyst was added. Another treatmentunder the same hydrogenation conditions completed the reaction. Then thecatalyst was removed by filtration and the crude product was purified bypreparative HPLC to give 97 mg of the Boc protected intermediate.

The protecting group was removed by treatment with 5-6 N HCl inisopropanol for 2 h at room temperature. The solvent was distilled ofand water and acetonitrile were added. Freeze drying of the mixture gave53 mg of compound 35.

LCMS Method #1, retention time 0.71 min, detected mass 257.18 [M+H]⁺

The following example compound was synthesized according to this method:

5-Ethyl-6-(piperidin-4-yloxy)-isoquinoline trifluoroacetate (36)

using compound 19A as the starting material

LCMS Method #2, retention time 0.17 min, detected mass 257.21 [M+H]⁺

Phenyl-[6-(piperidin-4-yloxy)-isoquinolin-5-yl]-methanol hydrochloride(37)

At −78° C. 0.6 mL (0.98 mmol, 1.6 M in hexane) n-butyl lithium wereadded to a solution of 200 mg (0.49 mmol, 1 eq.)4-(5-Bromo-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (11) in 3 mL of THF. After 30 min 110 μL (115 mg, 1.08 mmol) ofbenzaldehyde were added and the mixture was allowed to warm to ambienttemperature. After 2 h of stirring at room temperature water and ethylacetate were added. The layers were separated and the organic layer waswashed with water and brine. After drying over Na₂SO₄ and evaporation ofthe solvent the remainder was subjected to preparative HPLC to yield theBoc protected intermediate.

The Boc group was removed by dissolving the intermediate in isopropanoland addition of 5-6 N HCl in isopropanol. The precipitated hydrochloridewas isolated by filtration to yield 5.2 mg of compound 37 (3%).

¹H-NMR (d₆-DMSO): δ=9.43 (1H, s), 8.50 (1H, br s), 8.40 (1H, br s), 8.30(3H, m), 7.87 (1H, d, J=9.2 Hz), 7.33 (2H, d, J=7.4 Hz), 7.28 (2H, t,J=7.4 Hz), 7.19 (1H, t, J=7.4 Hz), 6.74 (1H, s), 6.34 (1H, s), 5.10 (1H,m), 3.25 (2H, m), 3.15 (2H, m), 2.19 (2H, m), 1.93 (2H, m).

LCMS Method #1, retention time 0.80 min, detected mass 335.22 [M+H]⁺

The following example compound was also synthesized according to thismethod:

1-[6-(Piperidin-4-yloxy)-isoquinolin-5-yl]-ethanol hydrochloride (38)

LCMS Method #1, retention time 0.55 min, detected mass 273.2 [M+H]⁺

2,2,2-Trifluoro-N-[6-(piperidin-4-yloxy)-isoquinolin-5-yl]-acetamidetrifluoro-acetate (39)

62.8 mg of potassium carbonate (0.46 mmol, 4 eq.) and 10.7 μL ofmethanesulfonyl chloride (0.13 mmol, 1.2 eq.) were added to a solutionof 39 mg (0.11 mmol) of4-(5-Amino-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (18) (containing some compound 11) in 3 mL DMF. The reaction wasstirred for 4 h at room temperature. Then water and ethyl acetate wereadded. The mixture was filtered through Celite, washed with ethylacetate and concentrated to yield a single product. The N-Boc-protectedintermediate was deprotected by treatment with 2 mL 5-6 N HCl inisopropanol for 2 h at room temperature. Then the solvent was evaporatedand the product was isolated by preparative HPLC to yield 18.5 mg ofcompound 39.

LCMS Method #1, retention time 0.39 min, detected mass 340.15 [M+H]⁺

N-[6-(Piperidin-4-yloxy)-isoquinolin-5-yl]-acetamide trifluoro-acetate(40)

Under argon atmosphere 81.5 mg (0.2 mmol) of4-(5-bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11) and 14.2 mg of acetamide (0.24 mmol, 1.2 eq.) were added to asolution of 27 mg (0.28 mmol, 1.4 eq) of NaOtBu in 3 mL toluene. Afterstirring for 10 minutes at room temperature 9.1 mg (0.01 mmol, 0.05 eq)Pd₂(dba)₃ and 11.9 mg (0.04 mmol, 0.2 eq) of2-(dt-butylphosphino)biphenyl were added. The reaction was heated to120° C. for 2 h in a microwave reactor (CEM Discovery). Then water andethyl acetate were added. The mixture was filtered through Celite,washed with ethyl acetate and concentrated. The remainder was subjectedtwice to preparative HPLC to yield the N-Boc-protected intermediate. TheN-Boc-protected intermediate was deprotected by treatment with 2 mL 5-6N HCl in isopropanol for 2 h at room temperature. Then the solvent wasevaporated and the product was isolated by preparative HPLC to yield 2.5mg of compound 40.

LCMS Method #3, retention time 0.15 min, detected mass 286.15 [M+H]⁺

General procedure for Boc-deprotection of building blocks:

The corresponding N-Boc-protected compounds were treated with 5-6 N HClin isopropanol for 2 h at room temperature. The precipitatedhydrochlorides were isolated by filtration and dried. If necessary,additional purification by preparative HPLC was performed.

LCMS Detected Starting Method Retention mass No. Compound material #time [M + H]⁺ 41

11 2 0.57 307.13 42

19A 2 0.64 255.19 43

15 2 0.46 254.15 44

12 1 0.67 355.04 45

16 1 0.64 254.13 46

20 1 0.87 311.12 47

19 1 0.72 255.19 48

 8 1 0.52 263.10

General procedure for Suzuki-coupling with4-(5-Bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11)

Aqueous Na₂CO₃ solution (0.2 ml, 0.4 mmol, 2 eq. 2M) was added to asolution of 81 mg (0.2 mmol, 1 eq.) of4-(5-Bromo-isoquinoline-6-yloxy)-piperidin-1-carboxylic acid tert-butylester (11) and 1.5 eq. (0.3 mmol) of the corresponding boronic acid(reagent 2) in 3 mL of DME. Argon was bubbled through the reactionmixture for 10 min. Then 23 mg (0.1 eq.) Pd(PPh₃)₄ were added and thereaction was stirred at 95° C. overnight under Argon atmosphere. Aftercooling 2 mL of water and 10 mL of ethyl acetate were added. The organiclayer was separated, dried and the solvent was distilled off. Theremainder was subjected to preparative HPLC.

The Boc group was removed by dissolving the intermediate in isopropanoland addition of 5-6 N HCl in isopropanol. The precipitate was isolatedby filtration.

In some reactions no hydrochloride precipitated or the purity of theprecipitate was unsatisfactory. In these cases the solvent was distilledoff and the remainder was purified by preparative HPLC.

The following examples were synthesized using this method:

Method retention detected mass No. Compound Reagent 1 Reagent 2 # time[M + H]⁺ 49

11

2 0.15 306.22 50

11

4 0.75 335.20 51

11

4 0.76 305.15 52

11

1 0.81 347.18 53

11

1 0.84 347.17 54

11

1 0.90 349.24 55

11

1 0.75 362.22 56

11

1 0.88 323.20 57

11

3 0.44 371.24 [M + MeCN + H]⁺ 58

11

3 1.09 389.13 59

11

3 1.02 345.14 60

11

3 1.06 373.03 61

11

1 0.87 350.16 62

11

1 0.72 383.14 62A

11

1 0.79 283.18 63

11

2 0.18 0.47 356.21 356.23 64

11

2 1.31 387.30 65

11

2 0.73 383.21 66

11

1 0.90 364.15 67

11

1 0.76 335.17 68

11

3 0.42 376.18 69

11

2 0.61 348.23 70

11

1 0.61 348.21 71

11

4 0.75 398.15 72

11

2 0.64 350.24 73

11

2 0.93 349.24 74

11

2 1.22 399.23 75

11

2 0.75 363.23 76

11

2 0.87 349.25 77

11

2 0.81 348.20 78

11

2 1.00 333.25Suzuki Coupling Procedure for Variation in the 5- and 7-Position

The base was added to a solution of reagent 1 (typically 0.2 mmol) andreagent 2 in DME. Argon was bubbled through the reaction mixture for 10min. Then the catalyst was added and the reaction was stirred at refluxtemperature overnight under Argon atmosphere. After cooling 2 mL ofwater and 10 mL of ethyl acetate were added. The mixture was filteredthrough a celite cartridge. The solvents were removed by distillationand the remainder was subjected to preparative HPLC.

The isolated intermediate was deprotected by treatment with 2 mL 5-6 NHCl in isopropanol for 2 h at room temperature. The solvent wasdistilled of and the precipitate was isolated by filtration. In somereactions no hydrochloride precipitated or the purity of the precipitatewas unsatisfactory. In these cases the solvent was distilled off and theremainder was purified by preparative HPLC.

Using this method the following examples were prepared:

LCMS Detected Method Retention mass No. Compound Reagent 1 Reagent 2Catalyst Base # time [M + H]⁺ 79

12

0.05 eq. PdOAc₂ 0.1 eq. P(Cy)₃ 3.5 eq. K₃PO₄ 2 0.66 269.17 80

12

0.05 eq. PdOAc₂ 0.1 eq. P(Cy)₃ 3.5 eq. K₃PO₄ 1 0.81 271.19 81

13

0.15 eq. Pd(PPh₃)₄ 2 eq. K₂CO₃ 1 0.87 305.16 82

13

0.15 eq. Pd(PPh₃)₄ 2 eq. K₂CO₃ 1 0.85 269.2

5-Isopropenyl-6-(piperidin-4-yloxy)-isoquinoline (83)

85 mg (0.62 mmol) of K₂CO₃ and 70 mg (0.15 mmol) of4-[5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-isoquinoline-6-yloxy]-piperidin-1-carboxylicacid tert-butyl ester (14) were added to a solution of 22 mg (0.18 mmol)2-Bromo-propene in 2 mL of DMF. Under argon atmosphere 5.6 mg (0.05 eq)of Pd(dppf)Cl₂ were added and the mixture was heated to 100° C. for 16h. After cooling to room temperature water and dichloromethane wereadded. The mixture was filtered through a Celite cartridge. The solventswere removed by distillation and the remainder was subjected topreparative HPLC. The isolated intermediate was deprotected by treatmentwith 2 mL 5-6 N HCl in isopropanol for 2 h at room temperature. Thesolvent was distilled of and compound 83 was isolated by preparativeHPLC to give 3.2 mg as the trifluoroacetate.

LCMS Method #3, retention time 0.15 min, detected mass 269.15 [M+H]⁺

Using this method the following examples were prepared:

LCMS retention detected mass No. Compound Reagent Method # time [M + H]⁺84

3 0.14 353.16 85

3 0.41 311.22 86

3 0.56 323.15

6-Methoxy-4-(4,4,4-trifluoro-butyl)-isoquinoline (87)

2 g 6-Methoxy-isoquinoline were dissolved in 25 mL of drytetrahydrofuran. 12.56 mL of a 1 M solution of potassium triethylborohydrate were added dropwise. The solution was allowed to stir atroom temperature for 2 h, then 3.29 g of 4,4,4-Trifluoro-1-iodobutanewere added dropwise. The solution was allowed to stir overnight, then 32mL of 1M sodium hydroxide and 12 mL of sodium peroxide solution (35%)were added. Stirring was continued for another 3 hrs, then the solutionwas diluted with dichloromethane, extracted with water and brine and theorganic layer was dried over sodium sulfate and evaporated to dryness.Silica gel chromatography yields 1.03 g of the desired product.

LCMS Method #1, retention time 1.20 min, detected mass 270.06 [M+H]⁺

6-Hydroxy-4-(4,4,4-trifluoro-butyl)-isoquinoline (88)

1.02 g of 6-Methoxy-4-(4,4,4-trifluoro-butyl)-isoquinoline (87) weretreated with boron tribromide as described for the synthesis of compound1 to give 410 mg of the desired product 88. LCMS Method #1, retentiontime 1.04 min, detected mass 256.00 [M+H]⁺

4-[4-(4,4,4-Trifluoro-butyl)-isoquinolin-6-yloxy]-piperidine-1-carboxylicacid tert-butyl ester (89)

100 mg of compound 88, 118 mg of Boc-(4-hydroxy)piperidine and 416 mg ofDiphenyl-[4-[1H,1H,2H,2H-perfluorodecyl]phenyl]phosphine were dissolvedin 5 mL of dry tetrahydrofuran. 208 mg of Bis (1H, 2H, 2H, 3H,3H-perfluorononyl)-azodicarboxylate were added and the reaction wasallowed to stir overnight. The mixture was evaporated to dryness andfiltered over a 5 g Fluoro-Flash cartridge. The obtained crude productwas purified by preparative HPLC to yield 46 mg of the desired product.

LCMS Method #1, retention time 1.51 min, detected mass 439.13 [M+H]⁺

6-(Piperidin-4-yloxy)-4-(4,4,4-trifluoro-butyl)-isoquinoline (90)

42 mg of compound 89 were dissolved in 5M hydrochloric acid inisopropanol. The solution was stirred at room temperature for 2 hrs andanother 2 hrs at 40° C., evaporated to dryness and taken up in water andlyophilized three times to give 32 mg of the desired product as thehydrochloride salt. LCMS Method #1, retention time 0.98 min, detectedmass 338.16 [M+H]⁺

The following isoquinolines were synthesized in a similar fashion asdescribed for compound 90, using appropriate alkyl halides:

Detected No. Compound Weight Method RT [min] Mass [MH⁺] 91

242.14 1 0.56 243.13 92

318.17 1 0.84 319.17General procedure for reductive amination:

1.5 eq of aldehyde was dissolved in 1 mL of methanol and 50 mg ofcompound 124 and 27 mg of anhydrous sodium acetate, dissolved inmethanol, were added. 0.250 mL of a solution of 1M sodiumcyanoborohydride in THF was added. The reaction was allowed to runovernight, then the solution was filtered, evaporated to dryness and theresidue was taken up in ethyl acetate. The organic layer was extractedwith a solution of 5% sodium carbonate in water, then with 5% sodiumchloride in water. The organic layer was dried, evaporated to drynessand purified by RP chromatography.

This procedure was used to obtain compounds 93 to 123:

No. Compound RT Mass [MH]⁺ 93

0.13 257.23 94

0.53 271.26 95

0.79 285.27 96

0.92 299.29 97

0.74 285.27 98

1.22 327.30 99

0.89 370.29 100

1.25 409.34 101

1.06 347.29 102

0.89 319.26 103

0.99 347.32 104

0.96 347.29 105

0.60 283.15 106

0.74 337.26 107

0.95 337.24 108

1.01 333.28 109

1.04 369.29 110

1.14 369.30 111

0.58/0.74 325.23 112

1.00 353.22/355.23 113

0.91 353.23/355.23 114

0.88 353.24/355.24 115

0.87 364.20 116

0.92 364.26 117

1.10 347.31 118

0.77 299.28 119

1.13 361.31 120

1.07 399.30 121

0.86 367.27 122

1.15 447.22/449.68 123

1.13 434.16

All LCMS in this table were obtained using LCMS method #2.

General Procedure for the Reaction of Boc-Protected Aminoalcohols with6-Hydroxy Isoquinolines (Mitsunobu-Reaction):

AAV1:

To 500 mg (1.5 mmol) of triphenylphosphine (bound to polystyrene, 3mmol/g) and 10 ml of dichloromethane 0.195 mL (1.2 mmol) ofdiethylazodicarboxylate (or alternatively diisopropylazodicarboxylate)were added. The reaction mixture was allowed to shake for 10 min. andthen 0.14 mL of triethylamine, 145 mg of 6-hydroxyisoquinoline (7) (oran equivalent amount of a different suitable isoquinoline) (reagent 1)and 1 mmol of the desired, boc-protected aminoalcohol (reagent 2) wasadded. The reaction was shaken at room temperature until no furtherconversion could be observed by LCMS. For workup, the solution wasfiltered, the residue was washed with dichloromethane and the organiclayer was washed twice with 1N sodium hydroxide, twice with water andonce with brine, dried over magnesium sulfate and evaporated. The crudeproduct was purified by preparative HPLC to yield the boc protectedcoupled product.

General Procedure for Removal of the Boc-Group (AAV2):

The starting material was dissolved in 2M hydrochloric acid and reactedovernight. To compounds with poor aqueous solubility, methanol ordioxane was added until a homogenous solution was obtained.Alternatively, 4M hydrochloric acid in isopropanol was used as thereactive compound. The reaction mixture was lyophilised and thedeprotected product is obtained as the corresponding hydrochloride ofthe free amine.

LCMS Retention Detected mass No. Compound Reagent 1 Reagent 2 Method #time [M + H]⁺ 124

7 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 1 0.47 229.21125

2 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.37 247.4126

2 3-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.34 247.35127

2 4-Piperidylmethanol-1- carboxylic acid tert-butyl ester 4 0.53 261.20128

2 3-Piperidylmethanol-1- carboxylic acid tert-butyl ester 4 0.57 261.20129

3 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.35 247.10130

3 3-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.35 247.10131

4 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 3 0.14 243.12132

8 3-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.60 263.15133

6 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 4 0.67 257.15134

5 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 1 0.68 257.20135

8 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 1 0.52 263.10136

9 4-Hydroxymethyl-piperidine- 1-carboxylic acid tert-butyl ester 2 0.33321.14/323.15 137

9 3-Hydroxy-piperidine- 1-carboxylic acid tert-butyl ester 2 0.31307.12/309.12 138

9 N-Methyl-3-piperidylmethanol 1 0.75 335.12 139

1 4-Hydroxypiperidine-1- carboxylic acid tert-butyl ester 2 0.62 307.06140

7 26 1 0.60 243.22 141

7 25 1 0.60 243.19 142

7 25 1 0.60 243.29 143

7 26 3 0.12 243.13 144

7 (1-Benzyl-piperidin-4-yl)- methanol 2 0.17 333.2 145

7 3-Hydroxy-piperidine- 1-carboxylic acid tert-butyl ester 2 0.2 229.2146

7 4-(2-Hydroxy-ethyl)- piperidine- 1-carboxylic acid tert-butyl ester 20.18 257.2 147

7 1-Methyl-piperidin-4-ol 1 0.45 243.2 148

7 3-Hydroxymethyl-piperidine- 1-carboxylic acid tert-butyl ester 2 0.17243.2 149

7 4-Hydroxymethyl-piperidine- 1-carboxylic acid tert-butyl ester 2 0.17243.2Chromatographic Resolution of Compounds 140 and 143:

The N-Boc protected intermediate, obtained as an enantiomeric mixture ofcompound 140 and compound 143, was separated into the enantiomers on achiral column (Chiralcel OD-H/56 250×4.6 mm). The removal of theprotection group as the final step was performed as described in thegeneral procedure.

The absolute configuration of the stereo centers has not beendetermined.

Compound 140: earlier eluting Boc-protected intermediate;

Compound 143: later eluting Boc-protected intermediate

Chromatographic Resolution of Compounds 141 and 142:

The N-Boc protected intermediate, obtained as an enantiomeric mixture ofcompound 141 and compound 142, was separated into the enantiomers on achiral column (Chiralpak AD-H/40 250×4.6 mm). The removal of theprotection group as the final step was performed as described in thegeneral procedure.

The absolute configuration of the stereo centers has not beendetermined.

Compound 141: earlier eluting Boc-protected intermediate;

Compound 142: later eluting Boc-protected intermediate.

7-Chloro-6-fluoro-isoquinoline (150)

7-Chloro-6-fluoro-isoquinoline is obtained by the same reactionsequence, used for the synthesis of 6-Fluoro-isoquinoline (23), startingfrom 3-Chloro-4-fluoro-benzaldehyde. R_(t)=0.77 min (Method #2).Detected mass: 182.1/184.1 (M+H⁺).

5-Chloro-6-fluoro-isoquinoline-trifluoro acetate (151)

7.0 g (38.1 mmol) of 6-Fluoroisoquinoline (23) are dissolved in 60 mL ofconcentrated sulfuric acid. At 0° C. 10.18 g of N-Chlorosuccinimide areadded. After 1 h another 5.2 g of N-Chlorosuccinimide are added and thesolution is warmed to 50° C. Two more portions of 5.2 gN-Chlorosuccinimide are added successively and stirring is continued at50° C. until the reaction is complete. The reaction mixture is cooled toroom temperature, is poured on ice and adjusted to pH 10 by addition ofsodium hydroxide. The precipitate is filtered off, taken up indichloromethane and washed with aqueous sodium hydroxide. The organiclayer is dried over magnesium sulfate, evaporated and the crude productis purified by preparative HPLC to yield 4.04 g of5-Chlor-6-fluor-isoquinoline (151) as trifluoroacetate. R_(t)=0.97 min(Method #2). Detected mass: 182.0/184.0 (M+H⁺).

6-Fluoro-5-nitro-isoquinoline (152)

Under cooling 2.0 mL of concentrated nitric acid are added to 2.8 mL ofsulphuric acid. Subsequently 350 mg of 6-fluoroisoquinoline (23) areadded, the reaction is warmed up to room temperature and allowed to stirovernight. The reaction mixture is poured on ice, extracted withdichloromethane and adjusted to alkaline pH by addition of sodiumhydroxide. The aqueous layer is extracted again with dichloromethane.The dichloromethane layer is dried over magnesium sulfate and evaporatedto give 90 mg of 6-Fluoro-5-nitro-isoquinoline, which can be usedwithout further purification. R_(t)=1.03 min (Method #1). Detected mass:193.0 (M+H⁺).

4-(Isoquinolin-6-yloxy)-piperidine-1-carboxylic acid-tert-butylester(154)

7.49 g of 4-Hydroxy-piperidine-1-carboxylic acid-tert-butylester aredissolved in 20 mL of dry dimethyl acetamide. 1.49 g of sodium hydride(60%) are added. Then a solution of 3.65 g 6-Fluoroisoquinoline (23) isadded dropwise. The solution is heated at 80° C. for 2 hours, then thesolvent is removed and the residue is taken up in dichloromethane. Theorganic layer is extracted twice with water and then with brine, driedover magnesium sulfate and evaporated to dryness. The crude product ispurified by silica gel chromatography to yield 6.22 g of4-(Isoquinolin-6-yloxy)-piperidine-1-carbocyclic acid-tert-butylester.R_(t)=1.32 min (Method #1). Detected mass: 329.1 (M+H⁺).

6-(Piperidin-4-yloxy)-isoquinoline hydrochloride (124)

4-(Isoquinolin-6-yloxy)-piperidine-1-carboxylic acid-tert-butylester(154) is deprotected by the general procedure described in AAV2 to yieldthe title compound as HCl-salt. R_(t)=0.20 min (Method #2). Detectedmass: 229.1 (M+H⁺).

The following example was synthesized according to this method:

4-Chloro-6-(piperidin-4-yloxy)-isoquinoline hydrochloride (156)

using 4-chloro-6-fluoro-isoquinoline (24) as starting material

LCMS Method #2, retention time 0.56 min, detected mass 263.12 [M+H]⁺

6-(1-Pyrimidin-2-yl-piperidin-4-yloxy)-isoquinoline-hydrochloride (157)

150 mg of 6-(Piperidine-4-yloxy)-isoquinoline hydrochloride (124) aredissolved in 10 mL of dry pyridine. 177 mg of triethylamine and 69 mg of4-chloropyrimidine are added and the solution is stirred at 65° C. for 6hours. The reaction mixture is poured on brine and extracted three timeswith ethyl acetate. The combined organic layers are dried over magnesiumsulfate, evaporated to dryness and the crude product is purified bypreparative HPLC. The product is converted into the corresponding HClsalt by taking up the product in 20 mL of 1 N hydrochloric acid followedby lyophilization. Yield: 47 mg. R_(t)=1.05 min (Method #2). Detectedmass: 307.1 (M+H⁺).

6-[1-(4-Trifluoromethyl-pyrimidin-2-yl)-piperidin-4-yloxy]-isoquinolinehydrochloride (158)

75 mg of 6-(Piperidine-4-yloxy)-isoquinoline-Hydrochloride (124) aredissolved in 5 mL of dry pyridine and 5 mL of DMF. 55 mg2-Chlor-4-trifluoromethyl-pyrimidine are added and the solution isstirred at 60° C. for 3 hours. The solvents are removed in vacuo and theresidue is taken up in brine and extracted three times with ethylacetate. The combined organic layers are dried over magnesium sulfate,evaporated to dryness and the crude product is purified by preparativeHPLC. The product is converted into the corresponding HCl salt by takingup the product in 20 mL of 1 N hydrochloric acid followed bylyophilization. Yield: 29 mg. R_(t)=1.69 min (Method #2). Detected mass:375.1 (M+H⁺).

6-(1-Cyclopropyl-piperidin-4-yloxy)-isoquinoline-hydrochloride (159)

300 mg (1.13 mmol) 6-(Piperidine-4-yloxy)-isoquinoline hydrochloride(124) are dissolved in 10 mL of methanol. 202 mg of triethylamine,molecular sieves 4A, 600 mg of glacial acetic acid, 871 mg of(1-Ethoxy-cyclopropyloxy)-trimethyl-silane and 101 mg of sodiumcyanoborohydride are added successively and the reaction mixture isheated under reflux for 6 hours. The reaction mixture is cooled to roomtemperature, 6 mL of 2N sodium hydroxide are added and the reactionmixture is filtered. The filtrate is evaporated, the residue is taken upin dichloromethane, extracted with 2 N sodium hydroxide and brine, driedwith sodium sulfate, evaporated to dryness and the crude material ispurified by preparative HPLC. The product fractions are evaporated, theproduct is taken up in 2 N hydrochloric acid and lyophilized.

Yield: 60 mg. R_(t)=0.50 min (Method #1). Detected mass: 269.2 (M+H⁺).

4-(2-Oxy-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (160)

3.97 g (12.1 mmol) of 4-(Isoquinolin-6-yloxy)-piperidine-1-carboxylicacid tert-butyl ester (154) are dissolved in 100 ml of dichloromethaneand 4.47 g (18.1 mmol) of 3-chloro-perbenzoic acid (70%) are added atroom temperature. The reaction mixture is stirred for 1 h and thenwashed with saturated sodium bicarbonate-solution. The aqueous phase isseparated and extracted with dichloromethane. The combined organiclayers are dried over magnesium sulfate and evaporated, to yield 4.19 gof crude material, which can be used for further conversion withoutpurification. R_(t)=1.46 min (Method #1). Detected mass: 345.2 (M+H⁺).

1-Chloro-6-(piperidin-4-yloxy)-isoquinoline-hydrochloride (161)

3.5 g (10.16 mmol) 4-(2-Oxy-isoquinolin-6-yloxy)-piperidine-1-carboxylicacid tert-butyl ester (160) were dissolved in 250 ml of HCl-saturatedethanol at 50° C. The clear solution was concentrated i. vac. and theresidue was refluxed in 50 ml POCl₃. After 3 h the POCl₃ was removed i.vac. and the residue was taken up in H₂O. The pH was adjusted to 11, byadding sodium hydroxide and the aqueous solution was extracted twicewith dichloromethane. The combined organic layers were dried overmagnesium sulfate and evaporated to dryness. The residue was purified bypreparative HPLC, by which the title compound was obtained astrifluoroacetate. This was converted to the corresponding HCl-salt bydissolving the product in 2 N HCl, followed by lyophilization. Yield:950 mg. R_(t)=1.03 min (Method #1). Detected mass: 263.1/265.1 (M+H⁺).

4-(1-Chloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (162)

1.23 g (4.11 mmol) of 1-Chloro-6-(piperidin-4-yloxy)-isoquinolinehydrochloride (161) were dissolved in 50 ml of dichloromethane and 0.85ml (6.15 mmol) of triethylamine were added. At 0° C. a solution of 1.09g (5.0 mmol) of tert-butyl-carbonate in 10 ml dichloromethane was addeddropwise and the mixture was allowed to stand at room temperatureovernight. For working up, the mixture was washed twice with H₂O, driedover magnesium sulfate and evaporated, to yield 1.1 g of the desiredproduct, which could be used without further purification. R_(t)=1.86min (Method #4). Detected mass: 363.1/365.2 (M+H⁺).

4-(1-Methylamino-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (163)

154 mg (0.42 mmol) of4-(1-Chloro-isoquinolin-6-yloxy)-piperidine-1-carbocyclic acidtert-butyl ester (162) were heated in 15 ml of an aqueousmethylamine-solution (41%)) at 110° C. in a sealed tube. After 7 h thereaction mixture was evaporated and the residue was taken up insaturated sodium bicarbonate solution and extracted with ethyl acetate.The organic layer was separated, dried over magnesium sulfate and thesolvent was removed i. vac. The residue was purified by silica gelchromatography (ethyl acetate/methanol 5:1). Yield: 45 mg. R_(t)=1.14min (Method #4). Detected mass: 358.3 (M+H⁺).

Methyl-[6-(piperidin-4-yloxy)-isoquinolin-1-yl]-amine-hydrochloride(164)

4-(1-Methylamino-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (163) was converted to the deprotected title compoundby the general procedure, described in AAV2, by which 34 mg of thecorresponding HCl-salt could be obtained. R_(t)=0.69 min (Method #1).Detected mass: 258.3 (M+H⁺).

Following the synthetic route, described for compound 164, the followingcompounds were prepared starting from4-(1-Chloro-isoquinolin-6-yloxy)-piperidine-1-carbocyclic acidtert-butyl ester (162) and the corresponding amines:

Mass Retention No. Compound Amine T [° C.] [MH⁺] time [min] Method 165

Ethylamine; 70% in H₂O 110° C. 272.28 0.72 1 166

Dimethylamine; 2M in THF 110° C. 272.29 0.68 1 167

Aniline; 1:1 in dioxan (v/v) 120° C. 320.20 0.88 1

2-Chloro-N-dimethylaminomethylene-5-formyl-benzenesulfonamide (168)

5.0 g (22.8 mmol) of 2-Chloro-5-formyl-benzenesulfonamide were dissolvedin 50 ml of dichloromethane. 4.08 g (34.3 mmol) of dimethylformamidedimethylacetal were added and the mixture was refluxed for 2 h. Aftercooling to room temperature, the solution was washed twice with H₂O,dried over magnesium sulfate and evaporated. 5.16 g of the crude productwere obtained and used in the next step without further purification.R_(t)=1.14 min (Method #1). Detected mass: 275.1/277.1 (M+H⁺).

2-Chloro-N-dimethylaminomethylene-5-[4-(isoquinolin-6-yloxy)-piperidin-1-ylmethyl]-benzenesulfonamide-trifluoroacetate (169)

200 mg (0.88 mmol) of 6-(piperidine-4-yloxy)-isoquinoline hydrochloride(124) were dissolved in 20 ml of methanol and 158 mg (1.56 mmol) oftriethylamine were added. After stirring for 15 minutes at roomtemperature 467 mg (7.78 mmol) of glacial acetic acid, 482 mg (1.76mmol) of 2-Chloro-N-dimethylaminomethylene-5-formyl-benzenesulfonamide(168), 166 mg (2.64 mmol) of sodium cyanoborohydride and freshly driedmolecular sieves were added and the mixture was refluxed for 3 h. Afterstirring overnight at room temperature, the mixture was filtered and thefiltrate was evaporated. The residue was dissolved in dichloromethaneand washed twice with saturated sodium bicarbonate solution and brine.After drying over magnesium sulfate and evaporation of the solvent, thecrude product was purified by preparative HPLC, by which 133 mg of thedesired product could be isolated as trifluoroacetate. R_(t)=0.87 min(Method #2), Detected mass: 487.2/489.2 (M+H⁺).

2-Chloro-5-[4-(isoquinolin-6-yloxy)-piperidin-1-ylmethyl]-benzenesulfonamide(170)

133 mg (0.27 mmol) of2-Chloro-N-dimethylaminomethylene-5-[4-(isoquinolin-6-yloxy)-piperidin-1-ylmethyl]-benzenesulfonamide(169) were dissolved in ethanol. After adding 50 ml of 2N NaOH, thesolution was heated to 60° C. for 6 h. After cooling to roomtemperature, the mixture was neutralized by addition of aqueous HCl andthe solvent was removed i. vac. The residue was stirred with ethanol,the inorganic salts were filtered off and the filtrate was evaporated.R_(t)=0.78 min (Method #2), Detected mass: 432.1 (M+H⁺).

4-(5-Nitro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (171)

90 mg (0.47 mmol) 6-Fluoro-5-nitro-isoquinoline (152) were treated with4-Hydroxy-piperidine-1-carboxylic acid tert-butyl ester following themethod described for the preparation of compound 154.

5-Nitro-6-(piperidin-4-yloxy)-isoquinoline-hydrochloride (172)

18.5 mg (0.05 mmol)4-(5-Nitro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acid tert-butylester (171) were deprotected following the procedure, described in AAV2,by which 12.5 mg of the title compound could be isolated as HCl-salt.R_(t)=0.57 min (Method #1). Detected mass: 274.2 (M+H⁺).

7-Chloro-6-(piperidin-4-yloxy)-isoquinoline-hydrochloride (173)

Starting from 7-Chloro-6-fluoro-isoquinoline (150), the title compoundwas prepared by the same synthetic route as for compound 124. R_(t)=0.66min (Method #1), detected mass: 263.1/265.1 (M+H⁺).

Synthetic procedure for the generation of 3,6-disubstitutedisoquinolines

Step 1

188 g of 5-Fluoro-indanone-1 (174) were dissolved in 1.81 of diethylether, 50 ml of EtOH saturated with HCl are added at 0° C. and 1.11 of a15% ethyl nitrite solution in ether is added over 1 hour.

The solution is allowed to stir for an additional 3 hours to reach roomtemperature, then the solvent is removed partially and the precipitatedproduct is collected by filtration.

Step 2

129 g of the product from Step 1 was added to a mixture of 170 g of PCl₅in 21 of POCl₃. Then gaseous HCl was added at 0° C. until saturation ofthe solution was reached. The remaining mixture was heated to 60° C. for6 h, the solvent partially removed in vacuo and the residue washydrolyzed on a crushed ice/water mixture. The precipitated product isisolated by filtration.

Step 3

155 g of product from Step 2 were added to a mixture of 740 ml HOAc and330 ml Hl (57%) containing 53 g of red phosphorous. After heating toreflux for 4 hours, the solution was treated with concentrated NaOH(until pH=8) and the precipitated product is isolated by filtration.

Step 4

16.5 g of N-Boc-4-hydroxypiperidine were dissolved in 210 ml of diglymeand treated with 4.1 g 50% NaH under nitrogen. The resulting mixture wasstirred for 1 h at room temperature, then 14.8 g of the product fromStep 4 was added. The mixture was allowed to stir for 1 day at roomtemperature, then 100 ml of toluene were added and the resulting mixturewas washed with water 3 times. The organic phases were collected and thesolvent was removed in vacuo.

Step 5

100 mg of compound 178 and 1.1 equivalents of the corresponding anilineare dissolved in 5 ml of dioxane, 350 mg of Cs₂CO₃, 20 mg of Pd(OAc)₂and 60 mg of XANTHPHOS are added and the resulting mixture is heated toreflux under nitrogen until the starting material is consumed. (reactionis monitored by LCMS) The solvent is removed in vacuo and the residue issubjected to chromatography on a HPLC system.

Step 6

The products of Step 5 are dissolved in 5 ml of ethanol saturated withgaseous HCl. After stirring for 5 h the desired product is isolated byremoval of the solvent in vacuo.

All 3,5,6-trisubstituted derivatives were synthesized according to theprocedure illustrated by the synthesis of compound 184. For synthesis ofcompound 185, acetamide was used as amine component in the Pd couplingstep.

Synthesis of4-(5-Bromo-3-chloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (181)

200 mg of 4-(3-Chloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (178) were dissolved in 5 ml of CH₃CN and heated to 85°C. Then a mixture of 148 mg of N-bromosuccinimide and 9 mg of AlBN wasadded as solid and the resulting mixture was heated to reflux for 1 h.The solvent was removed in vacuo and the residue subjected to flashcolumn chromatography. The yield of the isolated product was 41% LCMS:detected mass: 441.03, R_(t)=2.41 min (Method #1)

Synthesis of4-[3-Chloro-5-(4-fluoro-phenyl)-isoquinolin-6-yloxy]-piperidine-1-carboxylicacid tert-butyl ester (182)

150 mg of4-(5-Bromo-3-chloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (181) were dissolved in a mixture of 9 ml of dioxaneand 3 ml of water, 47 mg of 4-fluoro-benzene-boronic acid, 47 mg ofNa₂CO₃ and 40 mg of Pd(PPh₃)₄ were added and the resulting mixture washeated to 100° C. for 6 h. The solvent was removed in vacuo and theresidue subjected to chromatography on a HPLC system. Yield: 44% LCMS:detected mass: 457.22, R_(t)=2.45 min (Method #1)

Synthesis of4-[5-(4-Fluoro-phenyl)-3-(3,4,5-trimethoxy-phenylamino)-isoquinolin-6-yloxy]-piperidine-1-carboxylicacid tert-butyl ester (183)

70 mg of4-[3-Chloro-5-(4-fluoro-phenyl)-isoquinolin-6-yloxy]-piperidine-1-carboxylicacid tert-butyl ester(182) were dissolved in 7 ml of toluene, 20 mg ofPd(OAc)₂, 60 mg of XANTHPHOS, 400 mg of Cs₂CO₃ and 30 mg of 3,4,5trimethoxyaniline were added and the resulting mixture was heated to100° C. for 6 h. Then the solvent was removed in vacuo and the residuesubjected to chromatography on a HPLC system. The yield of isolatedproduct was 24%

LCMS: detected mass: 604.17, RT=1.81 min (Method #1)

Synthesis of[5-(4-Fluoro-phenyl)-6-(piperidin-4-yloxy)-isoquinolin-3-yl]-(3,4,5-trimethoxy-phenyl)-amine(184)

20 mg of4-[5-(4-Fluoro-phenyl)-3-(3,4,5-trimethoxy-phenylamino)-isoquinolin-6-yloxy]-piperidine-1-carboxylicacid tert-butyl ester (183) were dissolved in 5 ml of ethanol saturatedwith HCl (gaseous). The resulting mixture was stirred for 1 h, then thesolvent was evaporated and the product collected. Yield: 85%

LCMS: detected mass: 503.22, R_(t)=1.22 min (Method #1)

Detected No. Compound RT Method Mass [MH⁺] 185

0.12 #2 244.14 186

0.19 #2 286.18 187

0.17 #2 335.21 188

1.02 #2 335.21 189

1.02 #2 335.21 190

1.02 #2 335.21 191 1.02 #2 335.21 192

1.32 #2 335.20 193

1.21 #2 349.21 194

0.11 #2 329.34 195

0.17 #2 371.32 196

0.15 #2 316.21 197

1.18 #2 322.25 198

1..18 #2 312.18 199

1.29 #2 380.30 200

0.63 #2 385.29 201

0.2 #2 398.34 202

0.18 #2 369.30 203

0.2 #2 400.27 204

0.16 #2 455.27 205

0.16 #2 483.34 206

0.16 #2 434.26 206A

0.15 #2 452.30 207

0.15 #2 403.25 208

1.02 #2 321.17 209

1.02 #2 321.17 210

1.06 #2 321.17 211

0.85 #2 322.17 212

0.85 #2 322.17 213

0.89 #2 351.18 214

0.89 #2 351.18 215

1.03 #2 350.19 216

1 #2 380.20 217

1 #2 380.20 218

0.95 #2 335.19 219

1 #2 339.16 220

1.05 #1 380.20 221

1.06 #1 378.18 222

1.17 #1 378.18 223

1.12 #1 378.18 224

1.04 #1 408.19 225

1.04 #1 408.19 226

0.95 #1 378.18 227

1.1 #1 432.19 228

1.02 #1 410.21 230

1.12 #1 384.15 231

1.28 #1 412.20 232

1.31 #1 404.16 233

1.15 #1 433.22 234

1.25 #1 400.15 235

1.39 #1 442.21 236

1 #1 350.19 237

0.99 #1 350.19 238

1.13 #1 414.16 239

1.03 #1 364.20 240

1.5 #1 450.14 241

0.99 #1 380.20 242

1.25 #1 400.15 243

0.75 #1 315.18 244

0.83 #1 352.21 245

0.73 #1 322.17 246

0.84 #1 350.20 247

0.85 #1 335.19 248

1.05 #1 349.20 249

1.1 #1 348.21 250

1.1 #1 368.15 251

1.33 #1 420.14 252

1.12 #1 368.15 253

1.04 #1 354.14 254

1.07 #1 348.21 255

1.15 #1 456.23 256

1.11 #1 527.27 257

1.36 #1 540.21 258

1.15 #1 546.26 259

1.07 #1 486.24 260

1.08 #1 527.27 261

0.95 #1 364.17 262

1.08 #1 450.24 263

1.14 #1 486.24 264

1.03 #1 364.20 265

1.11 #1 390.22

4-Ethyl-6-(piperidin-4-yloxy)-isoquinoline (266)

The compound 266 was synthesized in a similar fashion as described forcompound 90, using ethyl iodide. LCMS Method #1, retention time 0.98min, detected mass 257.31 [M+H]⁺

Also using the same reaction sequence as for the synthesis of6-Fluoro-isoquinoline (23), the following two compounds were obtained:

8-Chloro-6-fluoro-isoquinoline (267)

R_(t)=0.83 min (Method #1). Detected mass: 182.12 (M+H⁺).

6-Fluoro-7-Methylisoquinoline (268)

R_(t)=0.70 min (Method #TOP). Detected mass: 162.3 (M+H⁺).

8-Chloro-6-(Piperidin-4-yloxy)-isoquinoline hydrochloride (269)

8-Chloro-6-(Piperidin-4-yloxy)-isoquinoline hydrochloride (269) wasobtained in a similar fashion as described above for the synthesis of(124), starting from 267 R_(t)=0.63 min (Method #1). Detected mass:263.14 (M+H⁺).

6-(Piperidin-4-yloxy)-7-Methyl isoquinoline hydrochloride (270)

6-(Piperidin-4-yloxy)-7-methyl isoquinoline hydrochloride (270) wasobtained in a similar fashion as described above for the synthesis of(124), starting from 268 R_(t)=0.64 min (Method #1). Detected mass:243.18 (M+H⁺).

The following set of compounds was obtained the same way, following thereductive amination procedure used to obtain examples 93-123 using 269,129 or 270, respectively and the corresponding aldehydes as startingmaterial. All LCMS in the following tables were obtained using LCMSmethod #2.

Example Mass No Formula T_(R) [MH⁺] 271

0.71 291.09 272

0.87 305.11 273

0.85 319.11 274

0.69 305.10 275

0.85 319.20 276

0.79 317.10 277

0.92 333.12 278

0.87 359.06 279

1.01 359.15 280

0.90 345.13 281

1.02 387.06 282

1.09 387.05 283

1.05 387.06 284

1.10 421.02 285

0.95 353.10 286

1.14 421.02 287

1.00 367.10 288

1.13 421.06 289

0.63 354.11 290

0.90 354.11 291

0.46 275.33 292

0.57 289.21 293

0.73 303.22 294

0.52 289.25 295

0.65 303.23 296

0.63 301.20 297

0.90 317.26 298

0.66 343.21 299

0.96 343.27 300

0.72 329.30 301

0.97 371.19 302

0.95 371.19 303

0.88 371.19 304

1.02 405.15/ 407.15 305

0.83 337.24 306

1.07 405.19/ 407.19 307

0.94 351.28 308

1.05 405.24 309

0.23 338.23 310

0.49 338.23 311

0.67 338.23 312

0.70 415.25 313

0.74 433.24 314

1.10 387.26 315

1.05 387.26 316

0.12 330.24 317

0.28 330.27 318

0.58 341.27 319

0.87 377.25 320

0.69 355.17 321

0.80 343.19 322

0.53 331.25 323

0.18 344.30 324

0.14 344.29 325

0.14 358.29 326

0.62 345.24 327

0.60 345.27 328

0.12 330.24 329

0.67 327.24 330

0.18 326.23 331

0.23 335.26 332

0.70 271.17 333

0.80 285.21 334

0.87 299.19 335

0.78 285.22 336

0.92 299.20 337

0.75 297.13 338

1.01 313.16 339

0.77 339.10 340

0.98 339.29 341

0.91 325.23 342

1.07 367.15 343

1.01 367.15 344

1.07 401.12/ 403.14 345

0.99 333.19 346

1.13 401.12/ 403.14 347

1.00 347.24 348

1.09 401.22 349

0.63 334.23 350

0.69 334.23 351

0.90 334.24 352

0.83 411.22 353

0.88 429.20 354

1.14 465.18 355

1.17 383.25 356

1.08 383.15 357

0.60 326.26 358

0.67 326.26 359

0.89 336.25 360

0.74 337.24 361

0.91 373.23 362

0.97 338.15 363

0.73 327.24 364

0.74 340.27 365

0.62 340.28 366

0.66 354.29 367

0.79 341.26 368

0.77 341.25 369

0.76 326.26 370

0.81 323.20 371

0.84 322.22 372

0.88 351.24

5,6,7-Trifluoro-isoquinoline (373)

5,6,7-Trifluoro-isoquinoline (373) is obtained by the same reactionsequence, used for the synthesis of 6-Fluoro-isoquinoline (23), startingfrom 3,4,5-Trifluorobenzaldehyde. Final purification by preparative HPLCgave the desired isoquinoline as trifluoroacetate. R_(t)=1.15 min(Method #2). Detected mass: 183.0.

4-(5,7-Difluoro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (374)

The title compound was synthesized following the protocol described for4-(Isoquinolin-6-yloxy)-piperidine-1-carbocyclic acid-tert-butylester(154). R_(t)=1.27 min (Method #TOP). Detected mass: 365.2 (M+H⁺).

5,7-Difluoro-6-(piperidin-4-yloxy)-isoquinoline (375)

4-(5,7-Difluoro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (374) is deprotected in Methanol/2 N HCl by the generalprocedure described in AAV2 to yield the title compound as HCl-salt.R_(t)=0.43 min (Method #TOP). Detected mass: 265.1 (M+H⁺).

5,7-Dichloro-6-fluoro-isoquinoline (376)

5.0 g (27.5 mmol) 7-Chloro-6-fluoro-isoquinoline (150) were dissolved in90 ml of conc. sulphuric acid. At room temperature 7.35 g (55.0 mmol)N-Chlorosuccinimide were added and the mixture was stirred at 50° C.After standing overnight at room temperature another 3 eq.N-Chlorosuccinimide were added and at the following again 5 eq.N-Chlorosuccinimide were added and the temperature was increased to 80°C. After no further conversion could be detected, the mixture was cooledto room temperature and poured on ice. The aqueous solution was broughtto basic pH by adding solid NaOH. The precipitate was filtered off andwashed three times with dichloromethane. After drying the organicfiltrates with MgSO₄ and evaporation of the solvent 1.09 g of thedesired isoquinoline could be isolated. R_(t)=1.26 min (Method #TOP).Detected mass: 216.0/218.0 (M+H⁺).

4-(5,7-Dichloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (377)

The title compound was synthesized following the protocol described for4-(Isoquinolin-6-yloxy)-piperidine-1-carbocyclic acid-tert-butylester(154). After final purification by preparative HPLC and evaporation ofthe product fractions, the Boc-group is already partially cleaved.R_(t)=1.71 min (Method #2). Detected mass: 397.2/399.2 (M+H⁺).

5,7-Dichloro-6-(piperidin-4-yloxy)-isoquinoline (378)

150 mg 4-(5,7-Dichloro-isoquinolin-6-yloxy)-piperidine-1-carboxylic acidtert-butyl ester (377, already partially deprotected) were dissolved in10 ml of dichloromethane and 1 ml of trifluoroacetic acid is added at 0°C. The solution is stirred for 2 h at room temperature. For working up,50 ml Dichloromethane were added and the solution was washed withsaturated NaHCO₃-solution. The layers were separated and the aqueousphase was extracted once with Dichloromethane. The combined organiclayers were again washed with saturated NaHCO3-solution, dried withMgSO4 and evaporated. The residue was purified by preparative HPLC. Theproduct fractions were evaporated and the residue dissolved in 2 N HCl.After evaporation, the title compound was isolated as HCl-salt.R_(t)=0.90 min (Method #2). Detected mass: 297.1/299.1 (M+H⁺).

Determination of Rho Kinase Inhibition

To measure Rho-kinase inhibition, IC₅₀ values were determined accordingto the following protocol:

Buffer: 25 mM Tris pH7.5; 0.02% BSA; 5% Glycerol; 0.008% Triton X100; 2%DMSO, 1 mM DTT; 1 mM MgCl₂; 0.5 μCi/well γ³³P ATP

Enzyme: ROCKII or ROKα) (Upstate, Catalog #14-451 Lot #24880U) 0.1 ng/μl

Final concentration of ATP in reaction mixture 40 μM

Biotinylated substrate, diluted to 0.25 μM with buffer described above(without ATP)

-   1. 10 μl Tris buffer (±Inhibitor)-   2. Add 30 μl of enzyme solution-   3. Start the reaction with 30 μL of mix substrate/ATP/ATP33-   4. Incubate for 20 min at room temperature-   5. Stop reaction with 30 μL of 50 mM EDTA-   6. Transfer 50 μL of stopped solution to Streptavidin Flash Plate    plus, Perkin Elmer, SMP 103A-   7. Incubate for 30 min at RT-   8. Wash 4 times with 300 μl of PBS/0.1% Tween 20-   9. Radioactivity in the well was determined

No. IC₅₀ 29 +++ 41 ++++ 44 ++++ 59 ++++ 67 +++ 72 ++++ 81 ++++ 111 +++100 +++ 120 ++++ 133 ++++ 134 +++ 138 ++++ 145 +++ 156 ++++ 228 ++++ 261++++ 265 +++ 266 +++ 269 ++++ 378 ++++

The given activity is denoted as the negative logarithm of the IC₅₀(pIC₅₀) as follows:

+: 3.0≦pIC₅₀<4.0

++ 4.0≦pIC₅₀<5.0

+++: 5.0≦pIC₅₀<6.0

++++: 6.0≦pIC₅₀

The invention claimed is:
 1. A method of treating Alzheimer's disease ina patient comprising administering to the patient at least one compoundof the formula (I):

wherein R₁ is H; R₂ is H; R₃ is H, NH-phenyl or NH-benzodioxolyl; R₄ isH, halogen, (C₁-C₆)alkyl; R₅ is H, halogen, (C₁-C₆)alkyl, cyclopropyl,phenyl, benzofuran, (C₁-C₆)alkylene-phenyl; R₆ is H, (C₁-C₈)alkyl,(C₁-C₆)alkylene-(C₁₀)aryl, (C₁-C₆)alkylene-CH[phenyl]₂,(C₁-C₆)alkylene-phenyl; R₇ is H, halogen, (C₁-C₆)alkyl, phenyl; R₈ is H,halogen, or (C₁-C₆)alkyl; n is 1, 2, 3 or 4; and L is O orO—(C₁-C₆)alkylene; wherein in residues R₄, R₅, R₇ and R₈ one alkyl oralkylene hydrogen atom can be optionally substituted by OH, OCH₃, COOH,COOCH₃, NH₂, NHCH₃, N(CH₃)₂, CONH₂, CONHCH₃ or CON(CH₃)₂, and in anyresidue one or more alkyl or alkylene hydrogen atoms can optionally besubstituted by F; and wherein phenyl or naphthyl groups are optionallysubstituted one or more times by (C₁-C₄)alkyl, O—(C₁-C₄)alkyl, O-phenyl,C(O)O—(C₁-C₆)alkyl, C(O)OH, C(O)—(C₁-C₄)alkyl, halogen, NO₂, SO₂NH₂, CN,SO₂—(C₁-C₄)alkyl, NH—SO₂—(C₁-C₄)alkyl, NH₂, NH—C(O)—(C₁-C₄)alkyl,(C₃-C₈)cycloalkyl, (C₁-C₄)alkyl-OH, C(O)N[(C₁-C₄)alkyl]₂, C(O)NH₂,N[(C₁-C₄)alkyl]₂, (C₂-C₄)alkenylene-(C₆-C₁₀)aryl, wherein the(C₆-C₁₀)aryl may be further substituted with (C₁-C₄)alkyl,(C₁-C₄)alkylene-O—(C₁-C₆)alkyl, O—(C₁-C₆)alkyl-(C₆-C₁₀)aryl, or may bevicinal substituted by a O—(C₁-C₄)alkylene-O group whereby a5-8-membered ring is formed together with the carbon atoms the oxygenatoms are attached to, and wherein the benzofuran or benzodioxolylgroups are optionally substituted one or more times with (C₁-C₄)alkyl,O—(C₁-C₄)alkyl, halogen or (C₁-C₄)alkylene-O—(C₁-C₄)alkyl, or apharmaceutically acceptable salt and physiologically functionalderivative thereof.
 2. A method according to claim 1, wherein in formula(I), R₃ is H, or NH-phenyl.
 3. A method according to claim 1, wherein informula (I), R₈ is H, halogen or (C₁-C₄)alkyl.
 4. A method according toclaim 3, wherein in formula (I), R₈ is H, Cl, F, methyl or ethyl.
 5. Amethod according to claim 1, wherein in formula (I), R₄ is H, halogen or(C₁-C₄)alkyl.
 6. A method according to claim 5, wherein in formula (I),R₄ is H.
 7. A method according to claim 1, wherein in formula (I), R₅ isH, halogen, (C₁-C₆)alkyl, phenyl, (C₁-C₂)alkylene-phenyl or benzofuran.8. A method according to claim 1, wherein in formula (I), R₇ is H,halogen, (C₁-C₄)alkyl, or phenyl.
 9. A method according to claim 1,wherein in formula (I), R₇ is H, fluoro, chloro, bromo, methyl, ethyl,or phenyl.
 10. A method according to claim 1, wherein in formula (I), nis 1, 2 or
 3. 11. A method according to claim 10, wherein in formula(I), n is
 1. 12. A method according to claim 1, wherein in formula (I),R₆ is H, (C₁-C₆)alkyl, (C₁-C₆)alkylene-(C₁₀)aryl or(C₁-C₆)alkylene-phenyl.
 13. A method according to claim 12, wherein informula (I), R₆ is H, (C₁-C₆)alkyl, (C₁-C₆)alkylene-(C₁₀)aryl or(C₁-C₆)alkylene-phenyl.
 14. A method according to claim 1, wherein informula (I), L is attached to the 3-position or to the 4-position of thepiperidine ring.
 15. A method according to claim 1, wherein in formula(I), L is attached to the 4-position of the piperidine ring.
 16. Amethod according to claim 1, wherein in formula (I), L is O-methylene,O-ethylene or O.
 17. A method according to claim 16, wherein in formula(I), L is O.
 18. A method according to claim 1, wherein in formula (I),R₁ is H; R₂ is hydrogen; R₃ is H, NH-phenyl or NH-benzodioxolyl; R₄ isH, halogen or (C₁-C₆)alkyl; R₅ is H, (C₁-C₆)alkyl, halogen, phenyl,(C₁-C₆)alkylene-phenyl or benzofuran; R₆ is H, (C₁-C₆)alkyl,(C₁-C₆)alkylene-(C₁₀)aryl or (C₁-C₆)alkylene-phenyl; R₇ is H, halogen,(C₁-C₆)alkyl, or phenyl; n is 1, 2 or 3, and L is O,O-methylene orO-ethylene.
 19. A method according to claim 1, wherein in formula (I),R₁ is H; R₂ is H; R₃ is H, halogen, or NH-phenyl or NH-benzodioxolyl; R₄is H, halogen or (C₁-C₄)alkyl; R₅ is H, (C₁-C₆)alkyl, halogen, phenyl,(C₁-C₆)alkylene-phenyl or benzofuran; R₆ is H, (C₁-C₆)alkyl,(C₁-C₆)alkylene-phenyl or (C₁-C₆)alkylene-(C₁₀)aryl; R₇ is H, halogen,(C₁-C₆)alkyl, or phenyl; n is 1, 2or 3; and L is O.
 20. A methodaccording to claim 1, wherein in formula (I), R₁ is H; R₂ is H; R₃ is Hor NH-phenyl; R₄ is H, halogen or (C₁-C₄)alkyl; R₅ is H, (C₁-C₄)alkyl,halogen, phenyl, or (C₁-C₂)alkylene-phenyl; R₆ is H, (C₁-C₆)alkyl,(C₁-C₆)alkylene-phenyl or (C₁-C₆)alkylene-(C₁₀)aryl; R₇ is H, halogen,(C₁-C₄)alkyl, or phenyl; R₈ is H, halogen or (C₁-C₄)alkyl; n is 1; and Lis O.
 21. A method according to claim 1 comprising administering to apatient a pharmaceutical composition comprising a pharmaceuticallyeffective amount of at least one compound of formula (I), or apharmaceutically acceptable salt thereof, a physiologically functionalderivative thereof and physiologically tolerated excipients andcarriers.
 22. A method according to claim 21 wherein the pharmaceuticalcomposition further comprises additional additives or other activeingredients.
 23. A method according to claim 1, wherein in formula (I),R₃ is


24. A method according to claim 1, wherein in formula (I), R₅ ishydrogen, fluoro, chloro, bromo, iodo, (p-methoxy)-phenyl, N-aniline,phenyl, benzyl, methyl, ethyl, cyclopropyl, 4-methoxy-anilin, N-acetylor a substituent of the group consisting of


25. A method according to claim 1 wherein in formula (I), R₆ is H,methyl, ethyl, propyl, butyl, s-butyl, pentyl, 3-methyl-butyl,isopropyl, benzyl or a substituent of the group consisting of